ZHOU XIAOYING (US)
CORREA MONA (US)
GATES DANIEL (US)
PARKER LEON (US)
FRANKLIN SCOTT (US)
| CLAIMS WHAT IS CLAIMED IS: 1. An oil comprising: at least 10 mg of ergosterol per 100 g of the oil; and a triacylglyceride (TAG) component, wherein at least 40% of TAGs in the TAG component are TAG species having a saturated fatty acid at the sn-2 position, wherein at least 50% of the acyl chains in the TAG component are C18:1. 2. The oil of claim 1, wherein the saturated fatty acid is C16:0. 3. The oil of any of the above claims, wherein at least 50% of the TAGs in the TAG component are TAG species having a saturated fatty acid at the sn-2 position. 4. The oil of any of the above claims, wherein the TAG species comprise C18:1 at the sn-1 and sn-3 positions. 5. The oil of any of the above claims, wherein the TAG species comprise C16:0 at the sn-2 position. 6. The oil of any of the above claims, wherein the TAG species comprises or consists of 1,3- dioleolyl-2-palmitoyl glycerol (OPO). 7. The oil of any of the above claims, wherein at least 20% of the acyl chains in the TAG component are C16:0. 8. The oil of any of the above claims, wherein at least 60% of the acyl chains in the TAG component are C18:1 and at least 30% of the acyl chains in the TAG component are C16:0. 9. The oil of any of the above claims, wherein 50-67% or 60-67% of the acyl chains in the TAG component are C18:1 and 20-33% of the acyl chains in the TAG component are C16:0. 10. The oil of any of the above claims, wherein the oil has an OP:OO m/z ratio of at least 1.6 as determined by abundance of diacylglycerol (DAG) ions resulting from mass spectrometry fragmentation of the TAG component in the oil. 11. The oil of any of the above claims, wherein the oil has an OP:OO m/z ratio of at least 2 as determined by abundance of DAG ions resulting from mass spectrometry fragmentation of the TAG component in the oil. 12. The oil of any of the above claims, wherein the oil has an OP:OO m/z ratio of at least 3 as determined by abundance of DAG ions resulting from mass spectrometry fragmentation of the TAG component in the oil. 13. The oil of any of the above claims, wherein the oil has an OP:OO m/z ratio of at least 4 as determined by abundance of DAG ions resulting from mass spectrometry fragmentation of the TAG component in the oil. 14. The oil of any of the above claims, wherein the oil comprises at least 50 mg of ergosterol per 100 g of the oil. 15. The oil of any of the above claims, wherein the oil comprises at least 100 mg of ergosterol per 100 g of the oil. 16. The oil of any of the above claims, wherein the oil comprises 100-200 mg of ergosterol per 100 g of the oil. 17. The oil of any of the above claims, wherein the oil comprises no more than 5 mg of campesterol, no more than 5 mg of β-sitosterol, or no more than 5 mg of stigmasterol per 100 g of the oil. 18. The oil of any of the above claims, wherein the oil comprises no more than 5 mg of campesterol per 100 g of the oil. 19. The oil of any of the above claims, wherein the oil does not comprise campesterol. 20. The oil of any of the above claims, wherein the oil comprises no more than 5 mg of stigmasterol per 100 g of the oil. 21. The oil of any of the above claims, wherein the oil does not comprise stigmasterol. 22. The oil of any of the above claims, wherein the oil comprises no more than 5 mg of β- sitosterol per 100 g of the oil. 23. The oil of any of the above claims, wherein the oil does not comprise β-sitosterol. 24. The oil of any of the above claims, wherein the oil further comprises one or more of ergosta-5,8-dien-3-ol, (3β)-, 5.xi.-ergost-7-en-3β-ol, 9,19-cyclolanostan-3-ol,24- methylene-,(3β)-, and ergosta-7,22-dien-3-ol, (3β). 25. The oil of claim 24, wherein the oil comprises at least 1 mg of ergosta-5,8-dien-3-ol, (3β)- per 100 g of the oil. 26. The oil of claim 24, wherein the oil comprises 1-50 mg of ergosta-5,8-dien-3-ol, (3β)- per 100 g of the oil. 27. The oil of claim 24, wherein the oil comprises at least 15 mg of 5.xi.-ergost-7-en-3β-ol per 100 g of the oil. 28. The oil of claim 24, wherein the oil comprises 1-50 mg of 5.xi.-ergost-7-en-3β-ol per 100 g of the oil. 29. The oil of claim 24, wherein the oil comprises at least 20 mg of 5.xi.-ergost-7-en-3β-ol per 100 g of the oil. 30. The oil of claim 24, wherein the oil comprises at least 5 mg of 9,19-cyclolanostan-3- ol,24-methylene-,(3β)- per 100 g of the oil. 31. The oil of any of the above claims, wherein the oil is an algal oil. 32. The oil of any of the above claims, wherein the oil is a genetically modified algal oil. 33. The oil of any of the above claims, wherein the oil is produced from an algal cell. 34. The oil of any of the above claims, wherein the oil is produced from a genetically modified algal cell. 35. The oil of any of the above claims, wherein the oil is produced from a Prototheca cell. 36. The oil of any of the above claims, wherein the oil is non-naturally occurring. 37. A composition comprising the oil of any one of claims 1-36; and one or more excipients. 38. The composition of claim 37, wherein the composition is a nutritional supplement. 39. The composition of claim 37, wherein the composition is an infant formula. 40. The composition of claim 39, wherein the infant formula comprises one or more of whey, casein, lactose, vitamin D, human milk oligosaccharides (HMOs), vegetable oil, and antibodies. 41. The composition of claim 40, wherein the vegetable oil comprises soy oil, canola oil, sunflower oil, coconut oil, palm oil, or palm kernel oil. 42. The composition of claim 39, wherein the infant formula comprises soy protein. 43. A TAG polyol produced from the oil of any one of claims 1-36. 44. A method for producing a TAG polyol, the method comprising: a) subjecting the oil of any one of claims 1-36 to epoxidation, thereby generating an epoxidized oil; and b) ring opening the epoxidized oil in the presence of an alcohol, an acid, or hydrogen and a suitable catalyst, thereby generating the TAG polyol. 45. The method of claim 44, wherein the epoxidized oil is ring opened in the presence of the alcohol. 46. The method of claim 44, wherein the epoxidized oil is ring opened in the presence of hydrogen and the suitable catalyst. 47. The method of claim 44, wherein the epoxidized oil is ring opened in the presence of the acid. 48. A method for producing a TAG polyol, the method comprising: a) subjecting the oil of any one of claims 1-36 to hydroformylation, thereby generating a hydroformylated oil; and b) reducing the hydroformylated oil in the presence of hydrogen and a suitable catalyst, thereby generating the TAG polyol. 49. The TAG polyol produced by the method of any one of claims 44-48. 50. A microalgal cell comprising an exogenous gene that encodes for an enzyme having lysophosphatidic acid acyltransferase activity, wherein the cell produces an oil comprising a TAG component, wherein at least 40% of TAGs in the TAG component are TAG species having a saturated fatty acid at the sn-2 position, wherein at least 50% of the acyl chains in the TAG component are C18:1. 51. The microalgal cell of claim 50, wherein the enzyme is a lysophosphatidic acid acyltransferase (LPAAT). 52. The microalgal cell of claim 50 or 51, wherein the enzyme is an algal lysophosphatidic acid acyltransferase (LPAAT). 53. The microalgal cell of any one of claims 50-52, wherein the enzyme comprises a sequence motif selected from any one of (I) NHXXXXD (or NHX4D); (II) (F/Y)XXR; (III) EGXR; and (IV) Proline, wherein X is any amino acid. 54. The microalgal cell of any one of claim 50-53, wherein the enzyme comprises NHXXXXD (or NHX4D), wherein X is any amino acid. 55. The microalgal cell of any one of claims 50-54, wherein the enzyme comprises a sequence with at least 70%, at least 85%, or 100% sequence identity to any one of SEQ ID NO: 107-109. 56. The microalgal cell of any one of claims 50-55, wherein the enzyme comprises (F/Y)XXR, wherein X is any amino acid. 57. The microalgal cell of any one of claims 50-56, wherein the enzyme comprises a sequence with at least 60%, at least 75%, at least 85%, or 100% sequence identity to any one of SEQ ID NO: 110-119. 58. The microalgal cell of any one of claims 50-57, wherein the enzyme comprises EGXR, wherein X is any amino acid. 59. The microalgal cell of any one of claims 50-58, wherein the enzyme comprises a sequence with at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 90%, or 100% sequence identity to any one of SEQ ID NO: 120-134. 60. The microalgal cell of any one of claims 50-59, wherein the enzyme comprises a sequence with a conserved proline. 61. The microalgal cell of any one of claims 50-60, wherein the enzyme comprises a sequence with at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 90%, or 100% sequence identity to any one of SEQ ID NO: 135-154. 62. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlamydomonas reinhardtii lysophosphatidic acid acyltransferase (CrLPAAT1). 63. The microalgal cell of any one of claims 50-52 and 62, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 12. 64. The microalgal cell of any one of claims 50-52, 62 and 63, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 2. 65. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlamydomonas incerta lysophosphatidic acid acyltransferase (CiLPAAT2). 66. The microalgal cell of any one of claims 50-52 and 65, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 15. 67. The microalgal cell of any one of claims 50-52, 65, and 66, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 10. 68. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlamydomonas schloesseri lysophosphatidic acid acyltransferase (ChsLPAAT2). 69. The microalgal cell of any one of claims 50-52 and 68, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 14. 70. The microalgal cell of any one of claims 50-52, 68, and 69, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 9. 71. The microalgal cell of any one of claims 50-52, wherein the enzyme is Volvox africanus lysophosphatidic acid acyltransferase (VaLPAAT2). 72. The microalgal cell of any one of claims 50-52 and 71, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 21. 73. The microalgal cell of any one of claims 50-52, 71, and 72, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 11. 74. The microalgal cell of any one of claims 50-52, wherein the enzyme is Nannochloropsis oceanica lysophosphatidic acid acyltransferase (NoLPAT3). 75. The microalgal cell of any one of claims 50-52 and 74, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 16. 76. The microalgal cell of any one of claims 50-52, 74, and 75, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 7. 77. The microalgal cell of any one of claims 50-52, wherein the enzyme is Synechocystis sp. 1-acyl-sn-glycerol-3-phosphate acyltransferase (Sll1848). 78. The microalgal cell of any one of claims 50-52 and 77, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 19. 79. The microalgal cell of any one of claims 50-52, 77, and 78, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 5. 80. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlamydomonas reinhardtii lysophosphatidic acid acyltransferase (CrLPAAT2). 81. The microalgal cell of any one of claims 50-52 and 80, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 13. 82. The microalgal cell of any one of claims 50-52, 80, and 81, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 1. 83. The microalgal cell of any one of claims 50-52, wherein the enzyme is Escherichia coli 1- acyl-sn-glycerol-3-phosphate acyltransferase (EcPlsC). 84. The microalgal cell of any one of claims 50-52 and 83, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 22. 85. The microalgal cell of any one of claims 50-52, 83, and 84, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 6. 86. The microalgal cell of any one of claims 50-52, wherein the enzyme is Volvox carteri lysophosphatidic acid acyltransferase (VcLPAAT2). 87. The microalgal cell of any one of claims 50-52 and 86, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 20. 88. The microalgal cell of any one of claims 50-52, 86, and 87, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 8. 89. The microalgal cell of any one of claims 50-52, wherein the enzyme is Brassica napus acyltransferase (BnBAT2). 90. The microalgal cell of any one of claims 50-52 and 89, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 18. 91. The microalgal cell of any one of claims 50-52, 89, and 90, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 3. 92. The microalgal cell of any one of claims 50-52, wherein the enzyme is Nannochloropsis oceanica lysophosphatidic acid acyltransferase (NoLPAT4). 93. The microalgal cell of any one of claims 50-52 and 92, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 17. 94. The microalgal cell of any one of claims 50-52, 92, and 93, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 4. 95. The microalgal cell of any one of claims 50-52, wherein the enzyme is Volvox carteri f. nagariensis lysophosphatidic acid acyltransferase (VcLPAAT2). 96. The microalgal cell of any one of claims 50-52 and 95, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 40. 97. The microalgal cell of any one of claims 50-52, 95, and 96, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 25. 98. The microalgal cell of any one of claims 50-52, wherein the enzyme is Astrephomene gubernaculifera lysophosphatidic acid acyltransferase (AgLPAAT2). 99. The microalgal cell of any one of claims 50-52 and 98, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 41. 100. The microalgal cell of any one of claims 50-52, 98, and 99, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 26. 101. The microalgal cell of any one of claims 50-52, wherein the enzyme is Edaphochlamys debaryana lysophosphatidic acid acyltransferase (EdLPAAT2). 102. The microalgal cell of any one of claims 50-52 and 101, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 42. 103. The microalgal cell of any one of claims 50-52, 101, and 102, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 27. 104. The microalgal cell of any one of claims 50-52, wherein the enzyme is Dunaliella salina lysophosphatidic acid acyltransferase (DsLPAAT2). 105. The microalgal cell of any one of claims 50-52 and 104, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 43. 106. The microalgal cell of any one of claims 50-52, 104, and 105, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 28. 107. The microalgal cell of any one of claims 50-52, wherein the enzyme is Scenedesmus sp. lysophosphatidic acid acyltransferase (SceLPAAT2). 108. The microalgal cell of any one of claims 50-52 and 107, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 44. 109. The microalgal cell of any one of claims 50-52, 107, and 108, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 29. 110. The microalgal cell of any one of claims 50-52, wherein the enzyme is Micractinium conductrix lysophosphatidic acid acyltransferase (McLPAAT2). 111. The microalgal cell of any one of claims 50-52 and 110, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 45. 112. The microalgal cell of any one of claims 50-52, 110 and 111, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 30. 113. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlorella sorokiniana lysophosphatidic acid acyltransferase (ChsoLPAAT2). 114. The microalgal cell of any one of claims 50-52 and 113, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 46. 115. The microalgal cell of any one of claims 50-52, 113, and 114, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 31. 116. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlorella variabilis lysophosphatidic acid acyltransferase (ChvaLPAAT2). 117. The microalgal cell of any one of claims 50-52 and 116, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 47. 118. The microalgal cell of any one of claims 50-52, 116, and 117, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 32. 119. The microalgal cell of any one of claims 50-52, wherein the enzyme is Raphidocelis subcapitata lysophosphatidic acid acyltransferase (RsLPAAT2). 120. The microalgal cell of any one of claims 50-52 and 119, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 48. 121. The microalgal cell of any one of claims 50-52, 119, and 120, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 33. 122. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlorella desiccata lysophosphatidic acid acyltransferase (ChdeLPAAT2). 123. The microalgal cell of any one of claims 50-52 and 122, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 49. 124. The microalgal cell of any one of claims 50-52, 122, and 123, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 34. 125. The microalgal cell of any one of claims 50-52, wherein the enzyme is Auxenochlorella protothecoides lysophosphatidic acid acyltransferase (ApLPAAT2). 126. The microalgal cell of any one of claims 50-52 and 125, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 50. 127. The microalgal cell of any one of claims 50-52, 125, and 126, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 35. 128. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chloropicon primus lysophosphatidic acid acyltransferase (ChprLPAAT2). 129. The microalgal cell of any one of claims 50-52 and 128, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 51. 130. The microalgal cell of any one of claims 50-52, 128, and 129, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 36. 131. The microalgal cell of any one of claims 50-52, wherein the enzyme is Homo sapiens lysophosphatidic acid acyltransferase (AGPAT1). 132. The microalgal cell of any one of claims 50-52 and 131, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 52. 133. The microalgal cell of any one of claims 50-52, 131, and 132, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 37. 134. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlamydomonas eustigma lysophosphatidic acid acyltransferase (CeLPAAT2). 135. The microalgal cell of any one of claims 50-52 and 134, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 53. 136. The microalgal cell of any one of claims 50-52, 134, and 135, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 38. 137. The microalgal cell of any one of claims 50-52, wherein the enzyme is Pedinophyceae lysophosphatidic acid acyltransferase (PedLPAAT2). 138. The microalgal cell of any one of claims 50-52 and 137, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 54. 139. The microalgal cell of any one of claims 50-52, 137, and 138, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 39. 140. The microalgal cell of any one of claims 50-52, 137, and 138, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 59. 141. The microalgal cell of any one of claims 50-52, 137, and 138, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 60. 142. The microalgal cell of any one of claims 50-52, 137, and 138, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 61. 143. The microalgal cell of any one of claims 50-52, 137, and 138, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 62. 144. The microalgal cell of any one of claims 50-52, wherein the enzyme is Volvox reticuliferus lysophosphatidic acid acyltransferase (VrLPAAT2). 145. The microalgal cell of any one of claims 50-52 and 144, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 55. 146. The microalgal cell of any one of claims 50-52, 144, and 145, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 57. 147. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlorella vulgaris lysophosphatidic acid acyltransferase (ChvuLPAAT2). 148. The microalgal cell of any one of claims 50-52 and 147, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 56. 149. The microalgal cell of any one of claims 50-52, 147, and 148, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 58. 150. The microalgal cell of any one of claims 50-52, wherein the enzyme is Volvulina compacta lysophosphatidic acid acyltransferase (VcomLPAAT2). 151. The microalgal cell of any one of claims 50-52 and 150, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 83. 152. The microalgal cell of any one of claims 50-52, 150, and 151, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 65. 153. The microalgal cell of any one of claims 50-52, wherein the enzyme is Vitreochlamys sp. CL-2021 lysophosphatidic acid acyltransferase (VitrLPAAT2). 154. The microalgal cell of any one of claims 50-52 and 153, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 84. 155. The microalgal cell of any one of claims 50-52, 153, and 154, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 66. 156. The microalgal cell of any one of claims 50-52, wherein the enzyme is Colemanosphaera charkowiensis lysophosphatidic acid acyltransferase (CchaLPAAT2). 157. The microalgal cell of any one of claims 50-52 and 156, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 85. 158. The microalgal cell of any one of claims 50-52, 156, and 157, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 67. 159. The microalgal cell of any one of claims 50-52, wherein the enzyme is Pleodorina japonica lysophosphatidic acid acyltransferase (PjapLPAAT2). 160. The microalgal cell of any one of claims 50-52 and 159, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 86. 161. The microalgal cell of any one of claims 50-52, 159, and 160, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 68. 162. The microalgal cell of any one of claims 50-52, wherein the enzyme is Volvulina boldii lysophosphatidic acid acyltransferase (VbolLPAAT2). 163. The microalgal cell of any one of claims 50-52 and 162, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 87. 164. The microalgal cell of any one of claims 50-52, 162, and 163, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 69. 165. The microalgal cell of any one of claims 50-52, wherein the enzyme is Pandorina morum lysophosphatidic acid acyltransferase (PmorLPAAT2). 166. The microalgal cell of any one of claims 50-52 and 165, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 88. 167. The microalgal cell of any one of claims 50-52, 165, and 166, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 70. 168. The microalgal cell of any one of claims 50-52, wherein the enzyme is Volvox carteri f. weismannia lysophosphatidic acid acyltransferase (VcarfLPAAT2). 169. The microalgal cell of any one of claims 50-52 and 168, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 89. 170. The microalgal cell of any one of claims 50-52, 168, and 169, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 71. 171. The microalgal cell of any one of claims 50-52, wherein the enzyme is Eudorina cylindrica lysophosphatidic acid acyltransferase (EcylLPAAT2). 172. The microalgal cell of any one of claims 50-52 and 171, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 90. 173. The microalgal cell of any one of claims 50-52, 171, and 172, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 72. 174. The microalgal cell of any one of claims 50-52, wherein the enzyme is Gonium multicoccum lysophosphatidic acid acyltransferase (GmulLPAAT2). 175. The microalgal cell of any one of claims 50-52 and 174, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 91. 176. The microalgal cell of any one of claims 50-52, 174, and 175, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 73. 177. The microalgal cell of any one of claims 50-52, wherein the enzyme is Gonium viridistellatum lysophosphatidic acid acyltransferase (GvirLPAAT2). 178. The microalgal cell of any one of claims 50-52 and 177, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 92. 179. The microalgal cell of any one of claims 50-52, 177, and 178, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 74. 180. The microalgal cell of any one of claims 50-52, wherein the enzyme is Volvox ferrisii lysophosphatidic acid acyltransferase (VferLPAAT2). 181. The microalgal cell of any one of claims 50-52 and 180, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 93. 182. The microalgal cell of any one of claims 50-52, 180, and 181, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 75. 183. The microalgal cell of any one of claims 50-52, wherein the enzyme is Vitreochlamys aulata lysophosphatidic acid acyltransferase (VaulLPAAT2). 184. The microalgal cell of any one of claims 50-52 and 183, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 94. 185. The microalgal cell of any one of claims 50-52, 183, and 184, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 76. 186. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlamydomonas sp. CCAC2762_B lysophosphatidic acid acyltransferase (Ch_CCAC2762_LPAAT2). 187. The microalgal cell of any one of claims 50-52 and 186, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 95. 188. The microalgal cell of any one of claims 50-52, 186, and 187, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 77. 189. The microalgal cell of any one of claims 50-52, wherein the enzyme is Dunaliella salina lysophosphatidic acid acyltransferase (DsalLPAAT2). 190. The microalgal cell of any one of claims 50-52 and 189, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 96. 191. The microalgal cell of any one of claims 50-52, 189, and 190, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 78. 192. The microalgal cell of any one of claims 50-52, wherein the enzyme is Microglena sp. YARC lysophosphatidic acid acyltransferase (MyarcLPAAT2). 193. The microalgal cell of any one of claims 50-52 and 192, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 97. 194. The microalgal cell of any one of claims 50-52, 192, and 193, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 79. 195. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlamydomonas sp. UWO_241 lysophosphatidic acid acyltransferase (CuwoLPAAT2). 196. The microalgal cell of any one of claims 50-52 and 195, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 98. 197. The microalgal cell of any one of claims 50-52, 195, and 196, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 80. 198. The microalgal cell of any one of claims 50-52, wherein the enzyme is Chlamydomonas moewusii lysophosphatidic acid acyltransferase (CmoeLPAAT2). 199. The microalgal cell of any one of claims 50-52 and 198, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 99. 200. The microalgal cell of any one of claims 50-52, 198, and 199, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 81. 201. The microalgal cell of any one of claims 50-52, wherein the enzyme is Oophila amblystomatis lysophosphatidic acid acyltransferase (OambLPAAT2). 202. The microalgal cell of any one of claims 50-52 and 201, wherein the enzyme comprises a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 100. 203. The microalgal cell of any one of claims 50-52, 201, and 202, wherein the cell comprises a sequence with at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 82. 204. The microalgal cell of any one of claims 50-203, wherein the exogenous gene is codon-optimized for expression in a Prototheca strain. 205. The microalgal cell of any one of claims 50-204, wherein the cell does not comprise an exogenous glycerol-3-phosphate acyltransferase (GPAT1). 206. The microalgal cell of any one of claims 50-205, wherein the cell is from a Prototheca base strain. 207. The microalgal cell of any one of claims 50-206, wherein the cell is from the Prototheca moriformis base strain UTEX 1533. 208. The microalgal cell of any one of claims 50-207, wherein the cell is derived from a classically improved strain from the Prototheca moriformis base strain UTEX 1533. 209. The microalgal cell of any one of claims 50-208, wherein the cell is from a non- genetically modified Prototheca base strain that produces an oil having a fatty acid profile of at least 50% oleic acid. 210. The microalgal cell of any one of claims 50-209, wherein the cell is from a non- genetically modified Prototheca base strain that produces an oil having a fatty acid profile of at least 30% palmitic acid. 211. The microalgal cell of any one of claims 50-210, wherein the Prototheca base strain is Prototheca wickerhamii. 212. The microalgal cell of any one of claims 50-210, wherein the Prototheca base strain is Prototheca moriformis. 213. The microalgal cell of any one of claims 50-212, wherein the microalgal cell produces at least 50% lipid by dry cell weight. 214. The microalgal cell of any one of claims 50-213, wherein the oil is the oil of any one of claims 1-36. 215. A method of producing a non-naturally occurring oil, the method comprising cultivating a microalgal cell in a culture medium, wherein the oil comprises a TAG component, wherein at least 40% of TAG species in the TAG component have a saturated fatty acid at the sn-2 position, wherein at least 50% of the acyl chains in the TAG component are C18:1. 216. The method of claim 215, further comprising isolating the oil composition from the culture medium. 217. The method of claim 215, further comprising expressing in the microalgal cell an exogenous enzyme having lysophosphatidic acid acyltransferase activity. 218. The method of any one of claims 215-217, wherein the microalgal cell is the microalgal cell of any one of claims 50-214. 219. The method of any one of claims 215-218, wherein the oil is the oil of any one of claims 1-36. |
[0141] In some embodiments, the oil contains a positive amount of ergosta-7,22-dien-3-ol, (3β,22E)-. In some embodiments, the amount of ergosta-7,22-dien-3-ol, (3β,22E)- in an oil provided herein is at least 2 mg, at least 5 mg, or at least 8 mg, or at least 9 mg per 100 g of the oil. In some embodiments, the amount of ergosta-7,22-dien-3-ol, (3β,22E)- in an oil is 1- 100 mg, 2-30 mg, 3-20 mg, or 5-15 mg per 100 g of the oil. [0142] In some embodiments, the oil contains a positive amount of ergosterol. In some embodiments, the amount of ergosterol in an oil provided herein is at least 50 mg, at least 100 mg, or at least 125 mg per 100 g of the oil. In some embodiments, the amount of ergosterol in an oil provided herein is at least 10 mg, at least 20 mg, at least 30 mg, at least 40 mg, at least 50 mg, at least 60 mg, at least 70 mg, at least 80 mg, at least 90 mg, at least 100 mg, at least 110 mg, at least 120 mg, at least 130 mg, at least 140 mg, at least 150 mg, at least 160 mg, at least 170 mg, at least 180 mg, at least 190 mg, or at least 200 mg per 100 g of the oil. In some embodiments, the amount of ergosterol in an oil is 10-50 mg, 50-100 mg, 100-150 mg, 150- 200 mg, 10-100 mg, 100-200 mg, 100-2000 mg, or 10-200 mg per 100 g of the oil. [0143] In some embodiments, the oil contains a positive amount of campesterol, a positive amount of β-sitosterol, or a positive amount of stigmasterol. In some embodiments, the amount of campesterol, β-sitosterol, or stigmasterol is no more than 5 mg per 100 g of the oil. In some embodiments, the oil does not contain campesterol. In some embodiments, the amount of campesterol in an oil provided herein is no more than 5 mg of campesterol per 100 g of the oil. In some embodiments, the oil does not contain stigmasterol. In some embodiments, the amount of stigmasterol in an oil is no more than 5 mg per 100 g of the oil. In some embodiments, the oil does not contain β-sitosterol. In some embodiments, the amount of β-sitosterol in an oil is no more than 5 mg per 100 g of the oil. [0144] In some embodiments, the oil contains a positive amount of ergosta-5,8-dien-3-ol, (3β)-. In some embodiments, the amount of ergosta-5,8-dien-3-ol, (3β)- is at least 1 mg per 100 g of the oil. In some embodiments, the amount of ergosta-5,8-dien-3-ol, (3β)- is at least 1 mg, at least 2 mg, at least 3 mg, at least 4 mg, at least 5 mg, at least 6 mg, at least 7 mg, at least 8 mg, at least 9 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 40 mg, at least 45 mg, or at least 50 mg per 100 g of the oil. In some embodiments, the amount of ergosta-5,8-dien-3-ol, (3β)- is 1-5 mg, 10-20 mg, 50-100 mg, 100-150 mg, 150-200 mg, 1-50 mg, 20-100 mg, 100-200 mg, or 20-200 mg per 100 g of the oil. [0145] In some embodiments, the oil contains a positive amount of 5.xi.-ergost-7-en-3β-ol. In some embodiments, the amount of 5.xi.-ergost-7-en-3β-ol is at least 15 mg per 100 g of the oil. In some embodiments the amount of 5.xi.-ergost-7-en-3β-ol is at least 20 mg per 100 g of the oil. In some embodiments, the amount of 5.xi.-ergost-7-en-3β-ol is at least 1 mg, at least 2 mg, at least 3 mg, at least 4 mg, at least 5 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 60 mg, at least 70 mg, at least 80 mg, at least 90 mg, at least 100 mg, at least 110 mg, at least 120 mg, at least 130 mg, at least 140 mg, at least 150 mg, at least 160 mg, at least 170 mg, at least 180 mg, at least 190 mg, or at least 200 mg per 100 g of the oil. In some embodiments, the amount of 5.xi.-ergost-7-en-3β-ol is 1-50 mg, 20-50 mg, 50-100 mg, 100- 150 mg, 150-200 mg, 20-100 mg, 100-2000 mg, or 20-200 mg per 100 g of the oil. [0146] In some embodiments, the oil contains a positive amount of 9,19-cyclolanostan-3- ol,24-methylene-,(3β)-. In some embodiments, the amount of 9,19-cyclolanostan-3-ol,24- methylene-,(3β)- is at least 5 mg of per 100 g of the oil. In some embodiments, the amount of 9,19-cyclolanostan-3-ol,24-methylene-,(3β)- is at least 1 mg, at least 2 mg, at least 3 mg, at least 4 mg, at least 5 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 40 mg, at least 45 mg, or at least 50 mg per 100 g of the oil. In some embodiments, the amount of 9,19-cyclolanostan-3-ol,24-methylene-,(3β)- is 5-10 mg, 10-20 mg, 20-30 mg, 30-40 mg, 40-50 mg, 5-25 mg, 25-50 mg, or 5-50 mg of per 100 g of the oil. [0147] TAG profile analysis of oil compositions provided herein can be conducted using liquid chromatography-mass spectrometry (LC-MS). TAG oils provided herein can be extracted from cells into a solution of 3:1 toluene/2-propanol (v/v) by means of mechanical disruption using ceramic beads and vigorous agitation. Filtered extracts can then be injected on LC mass spectrometer for profiling. [0148] Chromatographic separation of TAG regioisomers cannot be achieved by the LCMS method described. Instead, qualitative assessment of the regiospecificity of a predominant TAG species can be performed based on the abundance of diacylglycerol (DAG) ions resulting from fragmentation of the TAG species (FIG.1). In LC-APCI-MS analyses of TAGs, the loss of the fatty acid at the sn-1 and sn-3 positions are energetically more favorable compared to the sn-2 position, which results in a greater abundance of the DAG ions formed from those fragments. In the case of an OOP regioisomer, an approximate 1:1 ratio of the OP DAG ion (m/z 577) and OO DAG ion (m/z 603) can be observed, as shown in FIG.1, Panel A. In the case of the OPO regioisomer, a greater abundance of the OP DAG ion (m/z 577) can be observed, as shown in FIG.1, Panel B. [0149] FIG.1 demonstrates that TAG species of the same molecular weight show distinctive daughter ion ratios. The two TAG species, OOP and OPO in Panel A and Panel B, respectively, have identical mass/charge (m/z) ratios for the parent ion (859), but as illustrated, the molecules fragment very differently due to the effects of differing collision energies. Low energy collisions eliminate fatty acyl groups from the sn-1 or sn-3 position with greater frequency due to the relative inaccessibility of the acyl group at the sn-2 position. In the case of OOP, as shown in Panel A, low energy collisions result in daughter ions with m/z of 577 (OP) and 603 (OO), while less frequent high energy collisions resulting in the elimination of O from the sn-2 position also results in a daughter ion of m/z 577, OP. As a consequence, the resulting mass spectra of the daughter ions is the sum of all these fragments. The peak at 577 (OP) results from the more frequent elimination of O at sn-1 and the less frequent elimination of O at sn-2. Hence, the 577 fragment is roughly 1.5-2x the height of the 603 fragment (OO) resulting from the more frequent elimination of P at the sn-3 position. Alternatively, and as shown in Panel B, the OPO TAG species shows very different daughter ion ratios as the products of low energy collisions PO and OP have the same m/z of 577, while the daughter ion of the less frequently generated, high energy collision representing OO has a m/z of 603. For this TAG species, the 577 peak height, resulting from the summation of the low energy derived PO and OP daughter ions, is substantially greater than that for the 603 fragment, which requires much higher energy to generate. [0150] In some embodiments, the oil can have an OP:OO m/z ratio of as determined by abundance of DAG ions resulting from mass spectrometry fragmentation of the TAG component in the oil. In some embodiments, the oil can have an OP:OO m/z ratio of at least 1.6 as determined by abundance of DAG ions resulting from mass spectrometry fragmentation of the TAG component in the oil. In some embodiments, the oil can have an OP:OO m/z ratio of at least 1.6, at least 1.7, at least 1.8, at least 1.9, at least 2.0, at least 2.1, at least 2.2, at least 2.3 at least 2.4 at least 2.5, at least 2.6, at least 2.7, at least 2.8, at least 2.9, at least 3.0, at least 3.1, at least 3.2, at least 3.3, at least 3.4, at least 3.5, at least 3.6, at least 3.6, at least 3.7, at least 3.8, at least 3.9, at least 4.0, at least 4.1, at least 4.2, at least 4.3, at least 4.4, at least 4.5, at least 4.6, at least 4.7, at least 4.8, at least 4.9, or at least 5.0 as determined by abundance of DAG ions resulting from mass spectrometry fragmentation of the TAG component in the oil. [0151] TAG regiospecificity can further be determined by a porcine pancreatic lipase assay. This method can be used to determine identity of the sn-2 fatty acid of a TAG by incubating the TAG with porcine pancreatic lipase. The lipase reaction results in deacylation of the fatty acids at the sn-1 and sn-3 positions. The remaining sn-2-monoacylglycerols (MAGs) can then be isolated and identified by solid phase extraction and fatty acid composition analysis by gas chromatography using a direct transesterification method. [0152] Oil compositions provided herein can include a TAG component in which at least 40% of the TAGs in the TAG component are TAG species having a saturated fatty acid at the sn-2 position. In some embodiments, at least about 50% of the TAGs in the TAG component is a TAG species having a saturated fatty acid at the sn-2 position. In some embodiments, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, or about 60% of the TAGs in the TAG component is a TAG species having a saturated fatty acid at the sn-2 position. In some embodiments, 40-45%, 45-50%, 50-55%, 55-60%, 40-50%, 50-60%, or 40-60% of the TAGs in the TAG component is TAG species having a saturated fatty acid at the sn-2 position. In some embodiments, the saturated fatty acid at the sn-2 position is palmitate. [0153] Oil compositions provided herein can include a TAG component in which at least some of the acyl chains of the TAG component are C18:1. In some embodiments, at least about 50% of the acyl chains in the TAG component are C18:1. In some embodiments, at least about 50%, at least about 51%, at least about 52%, at least about 53%, at least about 54%, at least about 55%, at least about 56%, at least about 57%, at least about 58%, at least about 59%, at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, or at least about 70% of the acyl chains in the TAG component are C18:1. In some embodiments, at least about 50-55%, at least about 55-60%, at least about 60- 65%, at least about 60-67%, at least about 65-70%, at least about 50-60%, at least about 50- 67%, at least about 60-70%, or at least about 50-70% of the acyl chains in the TAG component are C18:1. Microbial Oils [0154] In some embodiments, an oil provided herein is obtained from a genetically modified microorganism, for example, microalgae, oleaginous yeast, or oleaginous bacteria. In some embodiments, the genetically modified microorganism is a genetically modified Prototheca sp. strain. In some embodiments, a genetically modified microorganism comprises an exogenous gene or exogenous nucleotides. Alternatively, or additionally, a genetically modified microorganism does not comprise an endogenous gene or endogenous nucleotides. Genetic modification methods described herein can be used to confer enriched oleic acid and/or OPO TAG production in a microorganism. [0155] An oil provided herein can be obtained from microalgae, oleaginous yeast, or oleaginous bacteria. In some embodiments, the microorganism is a modified Prototheca sp. strain. A non-genetically modified Prototheca sp. strain can be produced by classical strain improvement strategies. A genetically modified Prototheca sp. strain can be produced by expression of an exogenous gene and/or deletion of an endogenous gene in the microorganism. In some embodiments, a microorganism provided herein is a genetically modified Prototheca sp. strain produced by classical strain improvement strategies to optimize for enriched oleic acid production (e.g., at least 50% oleic acid) and enriched OPO production (e.g., at least 70% OPO). In some embodiments, a microorganism provided herein is a genetically modified Prototheca sp. strain produced by classical strain improvement strategies to optimize for enriched palmitic acid production (e.g., at least 30% palmitic acid) and enriched OPO production (e.g., at least 70% OPO). [0156] An oil provided herein can be produced from a Prototheca cell. The oil produced can be from a UTEX 1533 Prototheca strain cell. The oil produced can be from a cell of a strain that does not express an exogenous thioesterase. [0157] In some embodiments, an oil provided herein is produced by microalgae. In some embodiments, the microalgae is a species of a genus selected from the group consisting of: Chlorella sp., Pseudochlorella sp., Heterochlorella sp., Prototheca sp., Arthrospira sp., Euglena sp., Nannochloropsis sp., Phaeodactylum sp., Chlamydomonas sp., Scenedesmus sp., Ostreococcus sp., Selenastrum sp., Haematococcus sp., Nitzschia, Dunaliella, Navicula sp., Trebouxia sp., Pseudotrebouxia sp., Vavicula sp., Bracteococcus sp., Gomphonema sp., Watanabea, sp., Botryococcus sp., Tetraselmis sp., and Isochrysis sp. In some embodiments, the microalgae is Prototheca sp. In some embodiments, the microalgae is P. moriformis. In some embodiments, the microalgae is P. wickerhamii. Genetic Engineering and Classical Strain Improvement [0158] Production of TAGs oils having particular phenotypes can be achieved by genetic and non-genetic modification techniques of microorganisms. While genetic engineering techniques can be used to targeted to phenotypes elicited in a host oleaginous microbe, classical strain improvement and other non-genetic engineering techniques can be employed to further enhance these phenotypes. Similarly, classical strain improvement or other non- genetic engineering techniques can be employed to enhance certain phenotypes that have been selected by genetic engineering techniques. Phenotypes can include elaboration of a particular fatty acid profile, a particular TAG profile, yield on carbon, growth productivity, volumetric oil accumulation (e.g., g oil/L culture), oil productivity (e.g., g oil/L culture day), and oil as a percent dry cell weight (DCW) as a measure of strain performance. [0159] In some embodiments, a microalgal cell provided herein produces at least 50% lipid by dry cell weight. In some embodiments, a microalgal cell provided herein produces at least 60% lipid by dry cell weight. [0160] For example, a microorganism that has been non-genetically modified to confer a first phenotype (e.g., enriched oleic acid production) can be genetically modified to confer a second phenotype (e.g., enriched OPO production). The resulting microorganism can then confer both the first and second phenotype (e.g., enriched oleic acid and OPO production). [0161] Microorganisms provided herein can be produced by classical strain improvement strategies to select for microorganism having a desired phenotype, e.g., high oleic oil production. Classical strain improvement (also called “mutation breeding”) involves exposing organisms to chemicals or radiation to generate mutants with desirable traits. Ultraviolet (UV) light can be used to introduce random mutations within a microorganism’s nuclear genome. Chemical mutagens include compounds which inhibit or disrupt biosynthetic processes of a microorganism, e.g., antibiotics, antifungals, or carcinogens. Non-limiting examples of chemical mutagens include ICR-191, ethyl methanesulfonate (EMS), and 4- nitroquinoline-1-oxide (4-NQO). Non-limiting examples of chemical mutagens also include acridine mutagens, amino acid analogs, fatty acid biosynthesis inhibitors, cholesterol biosynthetic inhibitors, mTOR inhibitors, and membrane solubilizing agents. Combinations of chemical mutagens can also be used simultaneously to induce mutagenesis. Following mutagenesis, selective or enrichment agents can be used to select or enrich for strains of interest. Non-limiting examples of enrichment agents include L-canavanine, cerulenin, triparanol, clomiphene, clomiphene citrate, clotrimazole, terfenadine, fluphenazine, AZD8055, BASF 13-338, cafenstrole, clomiphene, PF-042110, and phenethyl alcohol. [0162] Microorganisms provided herein having enhanced or altered lysophosphatidic acid acyltransferase activity can be produced by genetic engineering techniques, such as genetic recombination. Enhancement of lysophosphatidic acid acyltransferase activity can be achieved by enhancing expression of an exogenous gene and/or reducing expression of an endogenous gene in the microorganism. Enhancement of gene expression can be through overexpression of an endogenous gene in a microorganism or expression of an exogenous gene. Reduction of gene expression can be through deletion or removal of an endogenous gene in a microorganism. [0163] For example, a non-naturally occurring microorganism provided herein can be genetically modified to enhance or alter lysophosphatidic acid acyltransferase activity in the microorganism. Enhancement or alteration of lysophosphatidic acid acyltransferase activity can be achieved by enhancing expression of an exogenous LPAAT gene and/or reducing expression of an endogenous LPAAT gene in the microorganism. [0164] Methods provided herein include classical strain improvement and/or genetic engineering methods to improve strain productivity, carbon yield, oleic acid content, and OPO content. Glucose consumption rate can be highly predictive indicator of lipid titer. As such, glucose consumption rate can be used as an enrichment tool in the mutant selection process. Polyol Applications and End Products [0165] Oils described herein can be used as substrates in chemistries used to produce polyols, for example hydroformylation/reduction or epoxidation and ring opening. Oleic acid moieties at the sn-1 and sn-3 position, when subjected to the aforementioned chemistries, will result in diols, that, when formulated with one or more excipients for a variety of applications, including but not limited to, process oils (e.g., for tires), waxes, lubricants, other polyols, macrodiols, other polyesterdiols, can be used to create polyurethane products, e.g., hard foams, soft foams, cast polyurethanes, thermoplastic polyurethanes (TPUs), elastomers, adhesives, coatings, laminates, films, and dispersions. Polyurethane products can be used to construct aerospace, automotive, medical, electronic, building and construction goods; sporting goods or recreational equipment, e.g., skis, snowboards, sidewalls, boating equipment, kayaks; and other consumer goods, e.g., industrial containers, coolers, mattresses, leather goods, apparel, footwear, mannequins, and phone cases. These polyurethane applications can serve as sustainable alternatives to petroleum-based, non-renewable materials, such as acrylonitrile butadiene styrene (ABS), ultra-high molecular weight polyethylene (UHMWPE), or high density polyethylene (HDPE). [0166] Oils provided herein can have improved production efficiency and a TAG composition that is enhanced for improved control of urethane chemistry. These characteristics of microbial oil can result in a greater degree of hydroxyl group (-OH) uniformity relative to oils with greater TAG heterogeneity (hence, lower purity) and/or diversity (e.g., oilseed or plant derived oils). Polyols derived from oils provided herein highly enriched in single, hydroxylated TAG species, can be preferable in generating polymers, including in instances where physical properties of a polymer can be compromised by molecular impurities, such as non-hydroxylated fatty acids or randomness in the regioselective insertion of fatty acid moieties on the glycerol backbone that may be present in oils having a more diverse or heterogeneous TAG profile. [0167] Polyols described herein can be particularly useful for producing polyurethane materials. For example, oils provided herein can have relatively low TAG diversity, low fatty acid diversity, and the fatty acids present in the oils may be hydroxylated fatty acids. A higher ratio of hydroxylated fatty acids to non-hydroxylated fatty acids can allow for increased chemical reactivity. Oils having low TAG diversity and a high proportion of hydroxylated fatty acids can be especially desirable in production of polyurethanes because hydroxylated fatty acids that can participate in crosslinking reactions with isocyanates. Thus, polyols generated from highly hydroxylated fatty acids comprising the oil, can yield polyurethane materials having superior properties. Polyol Production [0168] The hydroxyl group functionality of polyols can be introduced via a chemical conversion of a triglyceride oil. This conversion typically involves the presence of a double bond on an acyl moiety of the fatty acid, which can then be converted to incorporate one or more hydroxyl groups using several different chemistries including, for example, epoxidation/ring opening, ozonolysis, and hydroformylation and reduction. [0169] Epoxidation and subsequent ring opening across the carbon-carbon double bonds of an acyl chain can be carried out using a variety of reagents including, for example, water, hydrogen, methanol, ethanol, propanol, isopropanol, or polyols. Ring opening can be facilitated by reaction with an alcohol, including, for example, β-substituted alcohols. Ring opening of epoxidized oil can also be effected through the use of a hydrogenation catalyst, such as nickel, to create a hydroxyl moiety. [0170] Hydroformylation with synthesis gas (syngas) can be carried out using rhodium or cobalt catalysts to form the aldehyde at the olefinic group. The aldehyde can subsequently undergo reduction to an alcohol in the presence of hydrogen and a nickel catalyst to generate the polyol. [0171] The hydroformylation chemistry results in the preservation of fatty acid length and formation of primary hydroxyl group moieties. Primary hydroxyl group functionalities can be desirable in some PU applications due to increased reactivity compared to secondary hydroxyl group moieties. Hydroxyl groups introduced to olefinic groups in the acyl chain can participate in subsequent downstream chemistries, i.e., reaction with an isocyanate moiety to form a urethane linkage or reaction with methyl esters to form polyesters. Saturated fatty acids which do not contain double bonds cannot participate in crosslinking reactions with isocyanates. Hence, saturated fatty acids can compromise the structural integrity and degrade performance of the polymer produced therefrom. [0172] In some embodiments, a polyol provided herein contains a substantial proportion of primary hydroxyl groups. In some embodiments, a polyol provided herein contains secondary hydroxyl groups. In some embodiments, a polyol provided herein can be modified to increase the proportion of primary hydroxyl groups. [0173] Derivatives of TAG oils provided herein can be starting materials for producing polyols. Non-limiting examples of these TAG derivatives include fatty acids, fatty acid methyl esters, fatty acid ethyl esters, hydroxylated fatty acids, hydroxylated fatty methyl esters, and hydroxylated fatty ethyl esters. Non-limiting examples of polyols include polyester diols, polyether diols, hydrogenated polyols, hydroformylated polyols, and epoxidized, ring opened polyols. Alternatively, TAG oils provided herein, without further chemical modification, can be directly used as starting materials for generating polyols. [0174] Fatty acid methyl esters (FAMEs) can be generated through ester chemistry. For example, the TAG can be cleaved through transesterification into FAMEs and glycerol. In turn, FAMEs can be subjected to epoxidation and ring opening, for example, to create FAMEs of alcohols. Alternatively, polyols can first be generated from a TAG through epoxidation and ring opening, for example, followed by transesterification, into FAMEs of alcohols and glycerol. Glycerol and potassium methoxide catalyst can be removed by washing with water. [0175] Catalysts, including potassium methoxide (KOCH 3 ), 1,5,7-triazabicyclo[4.4.0]dec-5- ene (TBD), Titanium(IV) isopropoxide (TIP), dibutyltin dilaurate (DBTDL), tris(pentafluorophenyl)borane (BCF), and potassium tert-butoxide, among others, can be utilized to re-esterify ester groups to alcohol moieties. The dual functionality of alcohol FAMEs can be used to create polymer networks using only the methyl esters of the alcohol. Due to the polarity of the molecules (ester on one end and alcohol at the other end), the resulting polymer networks can elongate in a linear, unidirectional manner, and terminate in a single hydroxyl group. [0176] Polymer networks can also be elongated in a bi-directional manner by incorporation of a diol, such as low molecular weight diols or the polyols provided herein. Non-limiting examples of diols include propylene glycol, alkyl diols, 1,4-butanediol, 1,3-propanediol, and 1,6-hexanediol. In some embodiments, diols can be produced using microbial hosts. [0177] Hyperbranched polyols can be prepared to achieve a range of properties, such as molecular weight, viscosity, branching, and reactivity. For example, hyperbranched polyols can combine with isocyanates, ionogenic molecules, or hydrophobic compounds to produce higher order polyols. [0178] In some embodiments, TAG oils described herein can be utilized to create materials for polymer applications. As shown in FIG.7, TAG oils enriched in OPO can undergo chemical conversion to create oils containing functional groups, such as epoxy, ethoxy, methoxy, or hydroxy. Possible modifications of OPO include (from left to right): hydroformylation and reduction, resulting in a diol; epoxidation, resulting in the di-epoxy; and epoxidation followed by ring opening (e.g., with ethanol), resulting in a diol. TAGs enriched in OPO can allow for a variety of chemistries to be deployed around the carbon- carbon double bond of the oleic moieties, which can thereby produce numerous varieties of monomers for use as chain extenders in polymer chemistry applications. [0179] In some embodiments, strains producing OPO enriched TAGs such as those described herein can be useful substrates for additional genetic modifications with a delta-12 fatty acid desaturase (FAH12). Upregulation of FAH12 expression in such an OPO accumulating strain, can increase ricinoleic acid production, thereby increasing levels of ricinoleic acid incorporated into TAGs containing oleic acid. As a consequence, such strains can thereby produce diols, similar to those described in FIG.7, but in vivo without requiring additional chemical modifications. Non-limiting examples of oleate 12-hydroxylases include the Lesquerella fendleri FAH12, Ricinus communis FAH12, or Claviceps purpurea FAH12. An example of such a diol is shown in FIG.8. EXAMPLES Example 1: Expressing Chlamydomonas reinhardtii (CrLPAAT2) in P. moriformis strain CHK22. [0180] The microalgae Prototheca moriformis strain CHK22 (UTEX 1533) obtained from University of Texas at Austin Culture Collection of Algae produces a fatty acid ratio of oleate:palmitate of roughly 2:1, which can be suitable for OPO production. However, the P. moriformis lysophosphatidic acid acyltransferases (LPAATs) exhibit a strong preference for insertion of unsaturated fatty acids over saturated fatty acids at the sn-2 position. In this example, heterologous LPAAT genes were tested to identify genes that can efficiently direct palmitate (a saturated fatty acid) to the sn-2 position of TAGs produced by the microalgae. [0181] The C. reinhardtii CrLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK385 contained 5′ and 3′ homology arms to permit targeted integration of CrLPAAT2 into the genome and is shown in Table 1. Table 1: Integrative sequences for the transformation of P. moriformis with pCHK385 encoding CrLPAAT2.
[0182] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:CrLPAAT2:CvNR::3 ’DAO1b. Proceeding in the 5′ to 3′ direction, the bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination. The C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics, while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics. The P. moriformis PmAMT03 promoter, indicated by boxed, italicized text, drives the expression of the CrLPAAT2. The initiator ATG and terminator TGA codons of the CrLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0183] Strain Growth and Transformation [0184] Unless indicated otherwise, strains described herein were grown in 50-mL conical tube formats with shaking at 200 rpm at 28 ^C for 96 hours for lipid production. Culture supernatants were harvested via centrifugation and washed once with MilliQ water. The resulting cellular pellet was lyophilized to dryness and subjected to direct transesterification to generate fatty acid methyl esters (FAMEs) for subsequent quantitation and characterization by gas chromatography-flame ionization detection (GC/FID). [0185] Strains utilized for transformation were grown in vegetative growth medium for 24 hours prior to transformation. Cells were pelleted by centrifugation, resuspended in culture medium, and then re-centrifuged. The resulting cell pellet was re-suspended in culture medium and 5 × 10 7 cells plated to the appropriate selection medium and allowed to dry in a sterile biosafety cabinet. Transformants of CHK22 were generated via particle bombardment of previously plated cells using gold nanoparticles. Primary transformants were selected for ability to grow on plates containing sucrose as the sole carbon source. Transformants were grown in lipid production medium and their resulting biomass processed as previously described for fatty acid analysis. [0186] LCMS Analytical Method [0187] TAG profile analysis was conducted using LCMS. TAGs were extracted from cells into a solution of 3:1 toluene/2-propanol (v/v), by means of mechanical disruption using ceramic beads and vigorous agitation. The filtered extracts were then injected on an Agilent 1290 Infinity II UHPLC system coupled to a 6470B triple quadrupole mass spectrometer and APCI ionization source. [0188] LCMS Determination of Regioisomers [0189] Qualitative assessment of the regiospecificity of a predominant TAG species of the resulting oil was performed based on the abundance of DAG ions resulting from fragmentation of the TAG species (FIG.1). Example 2: Determination of the sn-2 fatty acid profile following digestion with pancreatic porcine lipase. [0190] The fatty acid composition at the sn-2 position was determined after incubating the TAGs with porcine pancreatic lipase. The lipase reaction results in deacylation of the TAG at the sn-1 and sn-3 positions, thereby leaving sn-2-MAGs. The sn-2-MAGs were then isolated using Agilent Bond Elut NH2 propyl solid phase extraction (SPE) cartridges and subjected to fatty acid composition analysis by gas chromatography using a direct transesterification method. [0191] Mass spectra and DAG ion ratios for the non-transgenic strain CHK22 and CHK22 transformed with pCHK385 (D552-1, D552-3, and D552-4) are shown in FIG.2 and Table 2. FIG.2 shows mass spectrum of OOP/OPO triglyceride. While regioisomers are not chromatographically separated by this analytical method, the higher ratio of m/z 577 (OP) to m/z 603 (OO) suggests a greater abundance of the OPO regioisomer compared to that of CHK22. Table 2: Screen of primary transformants of CHK22 with pCHK385. [0192] Table 2 shows the fatty acid/TAG profiles of four primary transformants of CHK22 transformed with pCHK385. P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization GC/FID. TAG analysis was conducted using the LCMS method as described above. [0193] The fatty acid composition at the sn-2 position of the transgenic lines CHK22, D552; 1, 3, and 4 were further determined after incubating the TAGs with porcine pancreatic lipase. The results were shown in Table 3. Table 3: Determination of the percentage of palmitate at sn-2 position following digestion with pancreatic porcine lipase, for the primary transformants of CHK22 with pCHK385. [0194] In Table 3, P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 120 hours, 200-mL culture in 1-L flask at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. The fatty acid composition at the sn-2 position was determined with the porcine pancreatic lipase assay as described above. A corresponding bar graph showing the percentage of palmitic acid at the sn-2 position for various strains that had been transformed with pCHK385 is shown in FIG.5. A graph showing LC-MS results for oil from CHK22 and strain D552-3 is shown in FIG.6, showing that D552-3 has a similar molecular signature to CHK22, but that the amounts of palmitic acid in the sn-2 position had significantly increased. Example 3: Expressing C. reinhardtii LPAAT1 (CrLPAAT1) in P. moriformis strain CHK22. [0195] C. reinhardtii CrLPAAT1 was introduced into a P. moriformis strain CHK22. The expression construct pCHK391 contained 5′ and 3′ homology arms to permit targeted integration of CrLPAAT1 into the genome and is shown in Table 4. Table 4: Integrative sequences for the transformation of P. moriformis with pCHK391 encoding CrLPAAT1.
[0196] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:CrLPAAT1:CvNR::3 ’DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed, uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′- UTR is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the CrLPAAT1. The initiator ATG and terminator TGA codons of the CrLPAAT1 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase, underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0197] Table 5 shows the fatty acid/TAG profiles of four primary transformants of CHK22 transformed with pCHK391. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 5: Screen of primary transformants of CHK22 with pCHK391. Example 4: Expressing Brassica napus BnBAT2 in P. moriformis strain CHK22. [0198] B. napus BnBAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK384 contained 5′ and 3′ homology arms to permit targeted integration of BnBAT2 into the genome and is shown in Table 6. Table 6: Integrative sequences for the transformation of P. moriformis with pCHK384 encoding BnBAT2.
[0199] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:BnBAT2:CvNR::3 DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the BnBAT2. The initiator ATG and terminator TGA codons of the BnBAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0200] Table 7 shows the fatty acid/TAG profiles of four primary transformants of CHK22 transformed with pCHK384. P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 7: Screen of primary transformants of CHK22 with pCHK384.
Example 5: Expressing Nannochloropsis NoLPAT4 in P. moriformis strain CHK22. [0201] Nannochloropsis NoLPAT4 was introduced into a P. moriformis strain CHK22. The expression construct pCHK386 contained 5′ and 3′ homology arms to permit targeted integration of NoLPAT4 into the genome and is shown in Table 8. Table 8: Integrative sequences for the transformation of P. moriformis with pCHK386 encoding NoLPAT4. [0202] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:NoLPAT4:CvNR::3 DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the NoLPAT4. The initiator ATG and terminator TGA codons of the NoLPAT4 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0203] Table 9 shows the fatty acid/TAG profiles of four primary transformants of CHK22 transformed with pCHK386. P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 9: Screen of primary transformants of CHK22 with pCHK386. Example 6: Expressing Synechocystis sp SII1848 in P. moriformis strain CHK22. [0204] Synechocystis sp Sll1848 was introduced into a P. moriformis strain CHK22. The expression construct pCHK387 contained 5′ and 3′ homology arms to permit targeted integration of Sll1848 into the genome and is shown in Table 10. Table 10: Integrative sequences for the transformation of P. moriformis with pCHK387 encoding Synechocystis sp SII1848.
[0205] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:Sll1848:CvNR::3 DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the Sll1848. The initiator ATG and terminator TGA codons of the Sll1848 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0206] Table 11 shows the fatty acid/TAG profiles of four primary transformants of CHK22 transformed with pCHK387. P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 11: Screen of primary transformants of CHK22 with pCHK387. Example 7: Expressing Escherichia coli EcPlsC in P. moriformis strain CHK22. [0207] The E. coli EcPlsC was introduced into a P. moriformis strain CHK22. The expression construct pCHK388 contained 5′ and 3′ homology arms to permit targeted integration of EcPlsC into the genome and is shown in Table 12. Table 12: Integrative sequences for the transformation of P. moriformis with pCHK388 encoding Escherichia coli EcPlsC. [0208] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:EcPlsC:CvNR::3 DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the EcPlsC. The initiator ATG and terminator TGA codons of the EcPlsC are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0209] Table 13 shows the fatty acid/TAG profiles of four primary transformants of CHK22 transformed with pCHK388. P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 13: Screen of primary transformants of CHK22 with pCHK388. Example 8: Expressing Nannochloropsis NoLPAT3 in P. moriformis strain CHK22. [0210] The Nannochloropsis NoLPAT3 was introduced into a P. moriformis strain CHK22. The expression construct pCHK453 contained 5′ and 3′ homology arms to permit targeted integration of NoLPAT3 into the genome and is shown in Table 14. Table 14: Integrative sequences for the transformation of P. moriformis with pCHK453 encoding Nannochloropsis NoLPAT3.
[0211] The construct can be written as 5’Thi4::PmHXT1v2:ScMEL1:PmPGK:CvNR:PmSAD2-2:NoLPAT3:CvNR:: 3’Thi4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from strain CHK22 that permit targeted integration at the Thi4 locus via homologous recombination. The P. moriformis hexose transporter 1 (HXT1 v2) promoter driving expression of the S. carlbergensis melibiase (ScMEL1) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for ScMEL1 are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. P. moriformis phosphoglucokinase (PGK) 3′-UTR is indicated by uppercase underlined text followed by a stearoyl ACP desaturase-2 (SAD2-2) promoter from P. moriformis, indicated by boxed italicized text, driving the expression of NoLPAT3. The initiator ATG and terminator TGA codons of NoLPAT3 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the P. moriformis Thi43′ flanking region indicated by bold, lowercase text. [0212] Table 15 shows the fatty acid/TAG profiles of four primary transformants of CHK22 transformed with pCHK453. P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 15: Screen of primary transformants of CHK22 with pCHK453. Example 9: Expressing Volvox carteri LPAAT2 (VcLPAAT2) in P. moriformis strain CHK22. [0213] V. carteri VcLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK785 contained 5′ and 3′ homology arms to permit targeted integration of VcLPAAT2 into the genome and is shown in Table 16. Table 16: Integrative sequences for the transformation of P. moriformis with pCHK785 encoding VcLPAAT2.
[0214] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:VcLPAAT2:CvNR::3 ’DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the VcLPAAT2. The initiator ATG and terminator TGA codons of the VcLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0215] Table 17 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK785. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 17: Screen of primary transformants of CHK22 with pCHK785. Example 10: Expressing Chlamdomonase schloesseri LPAAT2 (ChsLPAAT2) in P. moriformis strain CHK22. [0216] C. schloesseri ChsLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK786 contained 5′ and 3′ homology arms to permit targeted integration of ChsLPAAT2 into the genome and is shown in Table 18. Table 18: Integrative sequences for the transformation of P. moriformis with pCHK786 encoding ChsLPAAT2. [0217] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:ChsLPAAT2:CvNR:: 3’DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text followed by a linker. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the ChsLPAAT2. The initiator ATG and terminator TGA codons of the ChsLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0218] Table 19 shows the fatty acid/TAG profiles of six primary transformants of CHK22 transformed with pCHK786. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 19: Screen of primary transformants of CHK22 with pCHK786.
Example 11: Expressing Chlamydomonas incerta LPAAT2 (CiLPAAT2) in P. moriformis strain CHK22. [0219] C. incerta CiLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK787 contained 5′ and 3′ homology arms to permit targeted integration of CiLPAAT2 into the genome and is shown in Table 20. Table 20: Integrative sequences for the transformation of P. moriformis with pCHK787 encoding CiLPAAT2.
[0220] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:CiLPAAT2:CvNR::3 ’DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the CiLPAAT2. The initiator ATG and terminator TGA codons of the CiLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0221] Table 21 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK787. P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 21: Screen of primary transformants of CHK22 with pCHK787. Example 12: Expressing Volvox africanus LPAAT2 (VaLPAAT2) in P. moriformis strain CHK22. [0222] V. africanus VaLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK788 contained 5′ and 3′ homology arms to permit targeted integration of VaLPAAT2 into the genome and is shown in Table 22. Table 22: Integrative sequences for the transformation of P. moriformis with pCHK788 encoding VaLPAAT2.
[0223] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:VaLPAAT2:CvNR::3 ’DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the VaLPAAT2. The initiator ATG and terminator TGA codons of the VaLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The C. vulgaris nitrate reductase 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0224] Table 23 shows the fatty acid/TAG profiles of six primary transformants of CHK22 transformed with pCHK788. P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMES for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 23: Screen of primary transformants of CHK22 with pCHK788. [0225] As the transformants of CHK22 expressing CrLPAAT2 produced the higher OP:OO ratio, the percent identity (%) of all the amino acid sequences across the entire LPAAT protein was compared against that of CrLPAAT2. The results are shown in Table 24. The resulted phenotype of expressing these LPAATs in CHK22 is indicated with the m/z (577.5/603.5) OP:OO ratio; the OP:OO ratio shown in the rows of PmLPAAT1 or PmLPAAT2 represents the analytical results of the non-transgenic line CHK22. [0226] In some embodiments, a microalgal cell provided herein comprises an exogenous gene that encodes for a LPAAT in Table 24. In some embodiments, the exogenous gene can comprise a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a LPAAT in Table 24. Table 24: OP:OO Ratio [0227] Table 25 shows the amino acid sequences of LPAATs described in the Examples herein. FIG.3, Panels A-C illustrate amino acid sequence alignments of the amino acid sequences of LPAATs described herein, including EcPlsC (SEQ ID NO: 22), OlLPAAT2 (SEQ ID NO: 63), PmLPAAT2 (SEQ ID NO: 24), CosLPAAT2 (SEQ ID NO: 64), CrLPAAT2 (SEQ ID NO: 13), ChsLPAAT2 (SEQ ID NO: 14), CiLPAAT2 (SEQ ID NO: 15), VaLPAAT2 (SEQ ID NO: 21), VcLPAAT2 (SEQ ID NO: 20), NoLPAT4 (SEQ ID NO: 17), NoLPAT3 (SEQ ID NO: 16), PmLPAAT1 (SEQ ID NO: 23), Sll1848 (SEQ ID NO: 19), BnBAT2 (SEQ ID NO: 18), and CrLPAAT1 (SEQ ID NO: 12). [0228] In some embodiments, a microalgal cell provided herein comprises an exogenous gene that encodes for a LPAAT in Table 25. In some embodiments, the exogenous gene can comprise a sequence that encodes for an enzyme with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a LPAAT in Table 25. In some embodiments, the exogenous gene can comprise a sequence that encodes for an enzyme with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to any one of SEQ ID NO: 12-24, 63, and 64. Table 25: LPAAT Amino Acid Sequences Example 13: Expressing Volvox carteri f. nagariensis LPAAT2 (VcLPAAT2 V2) in P. moriformis strain CHK22. [0229] VcLPAAT2 V2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK967 contained 5′ and 3′ homology arms to permit targeted integration of VcLPAAT2 V2 into the genome and is shown in Table 27. Table 27: Integrative sequences for the transformation of P. moriformis with pCHK967 encoding VcLPAAT2 V2.
[0230] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:VcLPAAT2 V2:PmPGH::3’DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the VcLPAAT2 V2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the VcLPAAT2 V2. The initiator ATG and terminator TGA codons of the CrLPAAT2 V2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0231] Table 28 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK967. P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMES for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 28: Screen of primary transformants of CHK22 with pCHK967. Example 14: Expressing Astrephomene gubernaculifera LPAAT2 (AgLPAAT2) in P. moriformis strain CHK22. [0232] AgLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK968 contained 5′ and 3′ homology arms to permit targeted integration of AgLPAAT2 into the genome and is shown in Table 29. Table 29: Integrative sequences for the transformation of P. moriformis with pCHK968 encoding AgLPAAT2.
[0233] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:AgLPAAT2:PmPGH:: 3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the AgLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the AgLPAAT2. The initiator ATG and terminator TAG codons of the AgLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0234] Table 30 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK968. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 30: Screen of primary transformants of CHK22 with pCHK968. Example 15: Expressing Edaphochlamys debaryana LPAAT2 (EdLPAAT2) in P. moriformis strain CHK22. [0235] EdLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK969 contained 5′ and 3′ homology arms to permit targeted integration of EdLPAAT2 into the genome and is shown in Table 31. Table 31: Integrative sequences for the transformation of P. moriformis with pCHK969 encoding EdLPAAT2. [0236] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:EdLPAAT2:PmPGH:: 3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the EdLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the EdLPAAT2. The initiator ATG and terminator TAG codons of the EdLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0237] Table 32 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK969. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 32: Screen of primary transformants of CHK22 with pCHK969.
Example 16: Expressing Dunaliella salina LPAAT2 (DsLPAAT2) in P. moriformis strain CHK22. [0238] DsLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK971 contained 5′ and 3′ homology arms to permit targeted integration of DsLPAAT2 into the genome and is shown in Table 33. Table 33: Integrative sequences for the transformation of P. moriformis with pCHK971 encoding DsLPAAT2.
[0239] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:DsLPAAT2:PmPGH:: 3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the DsLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the DsLPAAT2. The initiator ATG and terminator TGA codons of the DsLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0240] Table 34 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK971. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 34: Screen of primary transformants of CHK22 with pCHK971. Example 17: Expressing Scenedesmus sp. LPAAT2 (SceLPAAT2) in P. moriformis strain CHK22. [0241] SceLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK973 contained 5′ and 3′ homology arms to permit targeted integration of SceLPAAT2 into the genome and is shown in Table 35. Table 35: Integrative sequences for the transformation of P. moriformis with pCHK973 encoding SceLPAAT2.
[0242] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:SceLPAAT2:PmPGH: :3’DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the SceLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the SceLPAAT2. The initiator ATG and terminator TGA codons of the SceLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0243] Table 36 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK973. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 36: Screen of primary transformants of CHK22 with pCHK973. Example 18: Expressing Micractinum conductrix LPAAT2 (McLPAAT2) in P. moriformis strain CHK22. [0244] McLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK974 contained 5′ and 3′ homology arms to permit targeted integration of McLPAAT2 into the genome and is shown in Table 37. Table 37: Integrative sequences for the transformation of P. moriformis with pCHK974 encoding McLPAAT2.
[0245] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:McLPAAT2:PmPGH:: 3’DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the McLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the McLPAAT2. The initiator ATG and terminator TGA codons of the McLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0246] Table 38 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK974. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 38: Screen of primary transformants of CHK22 with pCHK974. Example 19: Expressing Chlorella sorokiniana LPAAT2 (ChsoLPAAT2) in P. moriformis strain CHK22. [0247] ChsoLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK975 contained 5′ and 3′ homology arms to permit targeted integration of ChsoLPAAT2 into the genome and is shown in Table 39. Table 39: Integrative sequences for the transformation of P. moriformis with pCHK975 encoding ChsoLPAAT2. [0248] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:ChsoLPAAT2:PmPGH ::3’DAO1b. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the ChsoLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a Chlorella vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the ChsoLPAAT2. The initiator ATG and terminator TGA codons of the ChsoLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0249] Table 40 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK975. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 40: Screen of primary transformants of CHK22 with pCHK975. Example 20: Expressing Chlorella variabilis LPAAT2 (ChvaLPAAT2) in P. moriformis strain CHK22. [0250] ChvaLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK976 contained 5′ and 3′ homology arms to permit targeted integration of ChvaLPAAT2 into the genome and is shown in Table 41. Table 41: Integrative sequences for the transformation of P. moriformis with pCHK976 encoding ChvaLPAAT2.
[0251] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:ChvaLPAAT2:PmPGH ::3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the ChvaLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the ChvaLPAAT2. The initiator ATG and terminator TAG codons of the ChvaLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0252] Table 42 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK976. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 42: Screen of primary transformants of CHK22 with pCHK976. Example 21: Expressing Raphidocelis subcapitata LPAAT2 (RsLPAAT2) in P. moriformis strain CHK22. [0253] RsLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK977 contained 5′ and 3′ homology arms to permit targeted integration of RsLPAAT2 into the genome and is shown in Table 43. Table 44: Integrative sequences for the transformation of P. moriformis with pCHK977 encoding RsLPAAT2.
[0254] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:RsLPAAT2:PmPGH:: 3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the RsLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the RsLPAAT2. The initiator ATG and terminator TAG codons of the RsLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0255] Table 44 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK977. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 44: Screen of primary transformants of CHK22 with pCHK977. Example 22: Expressing Chlorella desiccata LPAAT2 (ChdeLPAAT2) in P. moriformis strain CHK22. [0256] ChdeLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK978 contained 5′ and 3′ homology arms to permit targeted integration of ChdeLPAAT2 into the genome and is shown in Table 45. Table 45: Integrative sequences for the transformation of P. moriformis with pCHK978 encoding ChdeLPAAT2.
[0257] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:ChdeLPAAT2:PmPGH ::3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the ChdeLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the ChdeLPAAT2. The initiator ATG and terminator TAG codons of the ChdeLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0258] Table 46 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK978. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 46: Screen of primary transformants of CHK22 with pCHK978. Example 23: Expressing Auxenochlorella proteothecoides LPAAT2 (ApLPAAT2) in P. moriformis strain CHK22. [0259] ApLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK979 contained 5′ and 3′ homology arms to permit targeted integration of ApLPAAT2 into the genome and is shown in Table 47. Table 47: Integrative sequences for the transformation of P. moriformis with pCHK979 encoding ApLPAAT2. [0260] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:ApLPAAT2:PmPGH:: 3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the ApLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the ApLPAAT2. The initiator ATG and terminator TAG codons of the ApLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0261] Table 48 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK979. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 48: Screen of primary transformants of CHK22 with pCHK979.
Example 24: Expressing Chloropicon primus LPAAT2 (ChprLPAAT2) in P. moriformis strain CHK22. [0262] ChprLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK980 contained 5′ and 3′ homology arms to permit targeted integration of ChprLPAAT2 into the genome and is shown in Table 49. Table 49: Integrative sequences for the transformation of P. moriformis with pCHK980 encoding ChprLPAAT2.
[0263] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:ChprLPAAT2:PmPGH ::3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the ChprLPAAT2gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the ChprLPAAT2. The initiator ATG and terminator TAG codons of the ChprLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0264] Table 50 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK980. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 50: Screen of primary transformants of CHK22 with pCHK980. Example 25: Expressing Homo sapiens AGPAT1 in P. moriformis strain CHK22. [0265] AGPAT1 was introduced into a P. moriformis strain CHK22. The expression construct pCHK966 contained 5′ and 3′ homology arms to permit targeted integration of AGPAT1 into the genome and is shown in Table 51. Table 51: Integrative sequences for the transformation of P. moriformis with pCHK966 encoding AGPAT1.
[0266] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:AGPAT1:PmPGH::3 DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the AGPAT1 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the AGPAT1. The initiator ATG and terminator TAG codons AGPAT1 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0267] Table 52 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK966. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 52: Screen of primary transformants of CHK22 with pCHK966. Example 26: Expressing Chlamydomonas eustigma LPAAT2 (CeLPAAT2) in P. moriformis strain CHK22. [0268] CeLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK970 contained 5′ and 3′ homology arms to permit targeted integration of CeLPAAT2 into the genome and is shown in Table 53. Table 53: Integrative sequences for the transformation of P. moriformis with pCHK970 encoding CeLPAAT2. [0269] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:CeLPAAT2:PmPGH:: 3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the CeLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drives the expression of the CeLPAAT2. The initiator ATG and terminator TAG codons of the CeLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0270] Table 54 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK970. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 54: Screen of primary transformants of CHK22 with pCHK970.
Example 27: Expressing Pedinophyceae sp. LPAAT2 (PedLPAAT2) in P. moriformis strain CHK22 using the PmAMT03 promoter. [0271] PedLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK972 contained 5′ and 3′ homology arms to permit targeted integration of PedLPAAT2 into the genome and is shown in Table 55. Table 55: Integrative sequences for the transformation of P. moriformis with pCHK972 encoding PedLPAAT2. [0272] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmAMT03:PedLPAAT2:PmPGH: :3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the PedLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmAMT03 promoter, indicated by boxed italicized text, drive the expression of the PedLPAAT2. The initiator ATG and terminator TAG codons of the PedLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0273] Table 56 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK972. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 56: Screen of primary transformants of CHK22 with pCHK972.
[0274] As the transformants of CHK22 expressing CrLPAAT2 produced the higher OP:OO ratio, the percent identity (%) of all the amino acid sequences across the entire LPAAT protein was compared against that of CrLPAAT2 the results are shown in Table 57. Pairwise identities (%) between all the sequences vs CrLPAAT2. The resulted phenotype of expressing these LPAATs in CHK22 is indicated with the m/z (577.5/603.5) OP:OO ratio; the OP:OO ratio shown in the rows of PmLPAAT1 or PmLPAAT2 represents the analytical results of the non-transgenic line CHK22. Amino acid sequences of the LPAATs described herein are listed in Table 58. Table 57: OP:OO Ratio
Table 58: LPAAT Amino Acid Sequences
Example 28: Expressing Volvox reticuliferus LPAAT2 (VrLPAAT2) in P. moriformis strain CHK22. [0275] VrLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1133 contained 5′ and 3′ homology arms to permit targeted integration of VrLPAAT2 into the genome and is shown in Table 59. Table 59: Integrative sequences for the transformation of P. moriformis with pCHK1133 encoding VrLPAAT2.
[0276] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmACP:VrLPAAT2:PmPGH::3 DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the VrLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. An acyl carrier protein promoter from P. moriformis (PmACP), indicated by boxed italicized text, drive the expression of the VrLPAAT2. The initiator ATG and terminator TAG codons of the VrLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0277] Table 60 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK1133. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 60: Screen of primary transformants of CHK22 with pCHK1133. Example 29: Expressing Chlorella vulgaris LPAAT2 (ChvuLPAAT2) in P. moriformis strain CHK22. [0278] ChvuLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1134 contained 5′ and 3′ homology arms to permit targeted integration of ChvuLPAAT2 into the genome and is shown in Table 61. Table 61: Integrative sequences for the transformation of P. moriformis with pCHK1134 encoding ChvuLPAAT2.
[0279] The construct can be written as 5’DAO1b::CrTUB2:ScSUC2:PmPGH:CvNR:PmACP: ChvuLPAAT2:PmPGH::3’DAO1b. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the DAO1b locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the ChvuLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmACP promoter, indicated by boxed italicized text, drives the expression of the ChvuLPAAT2. The initiator ATG and terminator TAG codons of the ChvuLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 DAO1b genomic region indicated by bold, lowercase text. [0280] Table 62 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK1134. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 62: Screen of primary transformants of CHK22 with pCHK1134. Example 30: Generation of a classically improved microalgal strain that produces a triglyceride oil enriched in palmitate. [0281] The microalgae P. moriformis strain CHK22 (UTEX 1533) was subjected to classical strain improvement to improve productivity, yield on carbon, and increase palmitic acid content. The mutagenesis, trait selection, and high-throughput, automated screening steps of the improvement process are outlined in the flow diagram in FIG.9. Briefly, algal cells in log phase of growth were subjected to mutagenesis by treatment with chemicals or UV light (1). Cells were then sub-cultured into lipid production medium and subjected to selection/enrichment strategies (2). Strains were then plated to solid medium to obtain clonal isolates (3), followed by the interrogation of these isolates in lipid production medium in a 96-well format (4). Using glucose consumption as a surrogate for oil production, high glucose consuming strains were validated in lipid production medium in a tube or shake flask format (5). Successfully validated isolates were sub-cultured for multiple generations to stabilize mutations (6), followed by purification of clonal isolates (7), and subsequent re- interrogation in lipid production medium (8). Phenotypically stable clones were then validated in fermentation (9). Clones showing variability in (8) were revalidated and sub- cultured via (6) to generate stable lines. [0282] CHK22 was chemically mutagenized for 30 min at 32 °C with 44 μM 4- nitroquinoline 1-oxide (4-NQO) or subjected to sham mutagenesis by the addition of only the mutagen solvent, DMSO. The mutagen was inactivated with the addition of sodium thiosulfate, which was then removed with repeated washing with water. The cells were then allowed to recover for 3 days in limited sugar growth media. The mutagenized and the mock- mutagenized populations were then independently cultured in lipid production media at 38 °C for five days. Optimal lipid production temperature for CHK22 is typically between 28-32 °C. A higher-than-optimal temperature was applied as a stressor to provide a growth advantage to mutants having a predisposition toward producing higher palmitate levels. At the end of the 5-day, 38 °C lipid culture, the cells were collected and then incubated at 65 °C for 4 min. Based on range-finding experiments conducted in the same format, exposure to temperatures within this range for this duration of time typically kills >99% of the cell population. After heat exposure, the cells were recovered in limited sugar growth media for three days, diluted, and then plated. Mutant clones were then selected and assessed for glucose consumption rate and fatty acid profile in a 72h, 96-well block-based lipid production assay (as illustrated in FIG.9). Table 63 summarizes glucose consumption rates and fatty acid profiles of TAG oil produced by the mutant strains. One mutant produced about 8% more palmitate than the CHK22 parent strain. This strain exhibited a significant reduction in glucose consumption rate, a hallmark of lower lipid titer. Table 63: Fatty acid profiles of select mutant strains exhibiting higher glucose consumption (Glc Rate) in a 96-well block lipid assay and increased palmitate (C16:0) levels. [0283] To further adapt the CHK22 mutant-3 strain to the altered fatty acid composition and improve lipid productivity, the strain was serially passaged in the absence of any selection for more than thirty population doublings and then plated to solid media to yield single colonies. These sub-clones were screened in a lipid production assay to assess the phenotypic stability of the lineage for both glucose consumption rate and fatty acid profile. After one cycle of serial passaging, the mutant was still phenotypically unstable. Several sub-clones that retained the higher palmitate level phenotype, along with increased rates of glucose consumption, were taken through additional rounds of serial passaging and stability assessment. After a total of five rounds of serial passaging and stability assessment, a stable strain, designated CHK100, was identified. CHK100 produced about 5% more palmitate than the CHK22 parental strain. The oil content (% on a weight-by-weight basis; %w/w), dry cell weight (DCW), and lipid titer (g/L) of CHK100 were also greater than those of CHK22 (Table 64 and Table 65). FIG.10 illustrates the strain improvement strategy of CHK22 to produce CHK100. Table 64: Tube-based assays on CHK100 showing improved lipid titer. NLB, non-lipid biomass; PCP, per cell production. Table 65: Tube-based assays on CHK100 showing increased C16:0 content.
[0284] All strains were subjected to standard lipid production conditions in which strains were grown in duplicate in 10 mL of lipid production medium. Cultures were grown with shaking (200 rpm) for 121 hours at 28 °C at which point about 1 mL of biomass was removed, applied to a polycarbonate filter, washed with an equal volume of Milli Q water, and placed in a tared glass vial at -80 °C for 30 minutes. Vials containing filters and frozen biomass were lyophilized to dryness overnight, weights were recorded, and filters with dried biomass were subjected to direct transesterification followed by GC/FID to quantitate FAMEs. Example 31: Expressing Pedinophyceae sp. LPAAT2 (PedLPAAT2) in P. moriformis strain CHK100 using the PmACP promoter. [0285] PedLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1126 contained 5′ and 3′ homology arms to permit targeted integration of PedLPAAT2 into the genome and is shown in Table 66. Table 66: Integrative sequences for the transformation of P. moriformis with pCHK1126 encoding PedLPAAT2. [0286] The construct can be written as 5’Thi4::CrTUB2:ScSUC2:PmPGH:CvNR:PmACP:PedLPAAT2:PmPGH::3 Thi4. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK100 that permit targeted integration at the Thi4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK100 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′- UTR is indicated by uppercase underlined text that enables amplification of the PedLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmACP promoter, indicated by boxed italicized text, drives the expression of the PedLPAAT2. The initiator ATG and terminator TAG codons of the PedLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK100 Thi4 genomic region indicated by bold, lowercase text. [0287] Table 67 shows the fatty acid/TAG profiles of primary transformants of CHK100 transformed with pCHK1126. The P. moriformis base strain CHK100 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 67: Screen of primary transformants of CHK100 with pCHK1126.
Example 32: Expressing Pedinophyceae sp. LPAAT2 (PedLPAAT2) in P. moriformis strain CHK100 using the PmG3PDH promoter. [0288] PedLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1127 contained 5′ and 3′ homology arms to permit targeted integration of PedLPAAT2 into the genome and is shown in Table 68. Table 68: Integrative sequences for the transformation of P. moriformis with pCHK1127 encoding PedLPAAT2.
[0289] The construct can be written as 5’Thi4::CrTUB2:ScSUC2:PmPGH:CvNR: PmG3PDH:PedLPAAT2:PmPGH::3’Thi4. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK100 that permit targeted integration at the Thi4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK100 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the PedLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drives the expression of the PedLPAAT2. The initiator ATG and terminator TAG codons of the PedLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK100 Thi4 genomic region indicated by bold, lowercase text. [0290] Table 69 shows the fatty acid/TAG profiles of primary transformants of CHK100 transformed with pCHK1127. The P. moriformis base strain CHK100 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 69: Screen of primary transformants of CHK100 with pCHK1127. Example 33: Expressing Pedinophyceae sp. LPAAT2 (PedLPAAT2) in P. moriformis strain CHK100 using the PmL40-2 promoter. [0291] PedLPAAT2 was introduced into a P. moriformis strain CHK100. The expression construct pCHK1128 contained 5′ and 3′ homology arms to permit targeted integration of PedLPAAT2 into the genome and is shown in Table 70. Table 70: Integrative sequences for the transformation of P. moriformis with pCHK1128 encoding PedLPAAT2.
[0292] The construct can be written as 5’Thi4::CrTUB2:ScSUC2:PmPGH:CvNR:PmL40- 2:PedLPAAT2:PmPGH::3’Thi4. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK100 that permit targeted integration at the Thi4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK100 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the PedLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmL40-2 promoter, indicated by boxed italicized text, drives the expression of the PedLPAAT2. The initiator ATG and terminator TAG codons of the PedLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK100 Thi4 genomic region indicated by bold, lowercase text. [0293] Table 71 shows the fatty acid/TAG profiles of primary transformants of CHK100 transformed with pCHK1128. The P. moriformis base strain CHK100 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 71: Screen of primary transformants of CHK100 with pCHK1128. Example 34: Expressing Pedinophyceae sp. LPAAT2 (PedLPAAT2) in P. moriformis strain CHK100 using the PmMPGp promoter. [0294] PedLPAAT2 was introduced into a P. moriformis strain CHK100. The expression construct pCHK1129 contained 5′ and 3′ homology arms to permit targeted integration of PedLPAAT2 into the genome and is shown in Table 72. Table 72: Integrative sequences for the transformation of P. moriformis with pCHK1129 encoding PedLPAAT2.
[0295] The construct can be written as 5’Thi4::CrTUB2:ScSUC2:PmPGH:CvNR: PmMPGp:PedLPAAT2:PmPGH::3’Thi4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK100 that permit targeted integration at the Thi4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK100 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR is indicated by uppercase underlined text that enables amplification of the PedLPAAT2 gene, followed by a linker, indicated by lowercase, bold italics, in which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmMPGp promoter, indicated by boxed italicized text, drives the expression of the PedLPAAT2. The initiator ATG and terminator TAG codons of the PedLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The P. moriformis PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK100 Thi4 genomic region indicated by bold, lowercase text. [0296] Table 73 shows the fatty acid/TAG profiles of primary transformants of CHK100 transformed with pCHK1129. The P. moriformis base strain CHK100 is shown as a non- transgenic control. Strains were grown for 96 hours in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 73: Screen of primary transformants of CHK100 with pCHK1129. Example 35: Expressing Volvulina compacta (VcomLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0297] VcomLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1210 contained 5′ and 3′ homology arms to permit targeted integration of VcomLPAAT2 into the genome and is shown in Table 74. Table 74: Integrative sequences for the transformation of P. moriformis with pCHK1210 encoding VcomLPAAT2.
[0298] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:VcomLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the VcomLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drives the expression of the VcomLPAAT2. The initiator ATG and terminator TAG codons of the VcomLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0299] Table 75 shows the fatty acid/TAG profiles of primary transformants of CHK22 transformed with pCHK1210. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 75: Screen of primary transformants of CHK22 with pCHK1210. Example 36: Expressing Vitreochlamys sp. CL-2021 (VitrLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0300] VitrLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1211 contained 5′ and 3′ homology arms to permit targeted integration of VitrLPAAT2 into the genome and is shown in Table 76. Table 76: Integrative sequences for the transformation of P. moriformis with pCHK1211 encoding VitrLPAAT2. [0301] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:VitrLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the VitrLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the VitrLPAAT2. The initiator ATG and terminator TGA codons of the VitrLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0302] Table 77 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1211. The P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 77: Screen of primary transformants of CHK22 with pCHK1211.
Example 37: Expressing Colemanosphaera charkowiensis (CchaLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0303] CchaLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1212 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 78. Table 78: Integrative sequences for the transformation of P. moriformis with pCHK1212 encoding CchaLPAAT2.
[0304] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:CchaLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the CchaLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the CchaLPAAT2. The initiator ATG and terminator TGA codons of the CchaLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0305] Table 79 shows the fatty acid/TAG profiles of eight primary transformants of CHK22 transformed with pCHK1212. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 79: Screen of primary transformants of CHK22 with pCHK1212. Example 38: Expressing Pleodorina japonica (PjapLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0306] PjapLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1213 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 80. Table 80: Integrative sequences for the transformation of P. moriformis with pCHK1213 encoding PjapLPAAT2.
[0307] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:PjapLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the PjapLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. A PmG3PDH promoter from P. moriformis, indicated by boxed italicized text, drive the expression of the PjapLPAAT2. The initiator ATG and terminator TGA codons of the PjapLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0308] Table 81 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1213. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 81: Screen of primary transformants of CHK22 with pCHK1213. Example 39: Expressing Volvulina boldii (VbolLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0309] VbolLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1214 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 82. Table 82: Integrative sequences for the transformation of P. moriformis with pCHK1214 encoding VbolLPAAT2. [0310] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:VbolLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the VbolLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the VbolLPAAT2. The initiator ATG and terminator TGA codons of the VbolLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0311] Table 83 shows the fatty acid/TAG profiles of eight primary transformants of CHK22 transformed with pCHK1214. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 83: Screen of primary transformants of CHK22 with pCHK1214. Example 40: Expressing Pandorina morum (PmorLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0312] PmorLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1215 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 84. Table 84: Integrative sequences for the transformation of P. moriformis with pCHK1215 encoding PmorLPAAT2.
[0313] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:PmorLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the PmorLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the PmorLPAAT2. The initiator ATG and terminator TGA codons of the PmorLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0314] Table 85 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1215. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 85: Screen of primary transformants of CHK22 with pCHK1215. Example 41: Expressing Volvox carteri f. weismannia (VcarfLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0315] VcarfLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1216 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 86. Table 86: Integrative sequences for the transformation of P. moriformis with pCHK1216 encoding VcarfLPAAT2. [0316] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:VcarfLPAAT2:PmPG H::3’Thia4. Proceeding in the 5′ to 3′ direction bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the VcarfLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the VcarfLPAAT2. The initiator ATG and terminator TGA codons of the VcarfLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0317] Table 87 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1216. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 87: Screen of primary transformants of CHK22 with pCHK1216. Example 42: Expressing Eudorina cylindrica (EcylLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0318] EcylLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1217 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 88. Table 88: Integrative sequences for the transformation of P. moriformis with pCHK1217 encoding EcylLPAAT2.
[0319] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:EcylLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the EcylLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the EcylLPAAT2. The initiator ATG and terminator TGA codons of the EcylLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0320] Table 89 shows the fatty acid/TAG profiles of eight primary transformants of CHK22 transformed with pCHK1217. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 89: Screen of primary transformants of CHK22 with pCHK1217. Example 43: Expressing Gonium multicoccum (GmulLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0321] GmulLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1218 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 90. Table 90: Integrative sequences for the transformation of P. moriformis with pCHK1218 encoding GmulLPAAT2.
[0322] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:GmulLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the GmulLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the GmulLPAAT2. The initiator ATG and terminator TGA codons of the GmulLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0323] Table 91 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1218. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 96 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 91: Screen of primary transformants of CHK22 with pCHK1218. Example 44: Expressing Gonium viridistellatum (GvirLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0324] GvirLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1219 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 92. Table 92: Integrative sequences for the transformation of P. moriformis with pCHK1219 encoding GvirLPAAT2.
[0325] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:GvirLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the GvirLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the GvirLPAAT2. The initiator ATG and terminator TGA codons of the GvirLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0326] Table 93 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1219. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 93: Screen of primary transformants of CHK22 with pCHK1219. Example 45: Expressing Volvox ferrisii (VferLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0327] VferLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1220 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 94. Table 94: Integrative sequences for the transformation of P. moriformis with pCHK1220 encoding VferLPAAT2.
[0328] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:VferLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the VferLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the VferLPAAT2. The initiator ATG and terminator TGA codons of the VferLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0329] Table 95 shows the fatty acid/TAG profiles of eight primary transformants of CHK22 transformed with pCHK1220. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 95: Screen of primary transformants of CHK22 with pCHK1220. Example 46: Expressing Vitreochlamys aulata (VaulLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0330] VaulLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1222 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 96. Table 96: Integrative sequences for the transformation of P. moriformis with pCHK1222 encoding VaulLPAAT2.
[0331] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:VaulLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the VaulLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the VaulLPAAT2. The initiator ATG and terminator TGA codons of the VaulLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0332] Table 97 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1222. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 97: Screen of primary transformants of CHK22 with pCHK1222.
Example 47: Expressing Chlamydomonas sp. CCAC2762_B (Ch_CCAC2762_LPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0333] Ch_CCAC2762_LPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1224 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 98. Table 98: Integrative sequences for the transformation of P. moriformis with pCHK1224 encoding Ch_CCAC2762_LPAAT2.
[0334] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:Ch_CCAC2762_LPAA T2:PmPGH::3’ Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the Ch_CCAC2762_LPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. A PmG3PDH promoter from P. moriformis, indicated by boxed italicized text, drive the expression of the Ch_CCAC2762_LPAAT2. The initiator ATG and terminator TGA codons of the Ch_CCAC2762_LPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0335] Table 99 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1224. The P. moriformis base strain CHK22 is shown as a non- transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 99: Screen of primary transformants of CHK22 with pCHK1224. Example 48: Expressing Dunaliella salina (DsalLPAAT2) in P. moriformis strain CHK22 using the PmG3PDG promoter. [0336] DsalLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1225 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 100. Table 100: Integrative sequences for the transformation of P. moriformis with pCHK1225 encoding DsalLPAAT2.
[0337] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:DsalLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the DsalLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. A PmG3PDG promoter from P. moriformis, indicated by boxed italicized text, drive the expression of the DsalLPAAT2. The initiator ATG and terminator TAG codons of the DsalLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic reg ion indicated by bold, lowercase text. [0338] Table 101 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1225. The P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 101: Screen of primary transformants of CHK22 with pCHK1225.
Example 49: Expressing Microglena sp. YARC (MyarcLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0339] MyarcLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1227 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 102. Table 102: Integrative sequences for the transformation of P. moriformis with pCHK1227 encoding MyarcLPAAT2.
[0340] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:MyarcLPAAT2:PmPG H::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the MyarcLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. A PmG3PDH promoter from P. moriformis, indicated by boxed italicized text, drive the expression of the MyarcLPAAT2. The initiator ATG and terminator TGA codons of the MyarcLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0341] Table 103 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1227. The P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 103: Screen of primary transformants of CHK22 with pCHK1227. Example 50: Expressing Chlamydomonas sp. UWO_241 (CuwoLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0342] CuwoLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1228 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 104. Table 104: Integrative sequences for the transformation of P. moriformis with pCHK1228 encoding CuwoLPAAT2. [0343] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:CuwoLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the CuwoLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the CuwoLPAAT2. The initiator ATG and terminator TGA codons of the CuwoLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0344] Table 105 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1228. The P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 105: Screen of primary transformants of CHK22 with pCHK1228.
Example 51: Expressing Chlamydomonas moewusii (CmoeLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0345] CmoeLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1231 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 106. Table 106: Integrative sequences for the transformation of P. moriformis with pCHK1231 encoding CmoeLPAAT2.
[0346] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:CmoeLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the CmoeLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter , indicated by boxed italicized text, drive the expression of the CmoeLPAAT2. The initiator ATG and terminator TGA codons of the CmoeLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0347] Table 107 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1231. The P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 107: Screen of primary transformants of CHK22 with pCHK1231. Example 52: Expressing Oophila amblystomatis (OambLPAAT2) in P. moriformis strain CHK22 using the PmG3PDH promoter. [0348] OambLPAAT2 was introduced into a P. moriformis strain CHK22. The expression construct pCHK1232 contained 5′ and 3′ homology arms to permit its targeted integration into the genome and is shown in Table 108. Table 108: Integrative sequences for the transformation of P. moriformis with pCHK1232 encoding OambLPAAT2.
[0349] The construct can be written as 5’Thia4::CrTUB2:ScSUC2:PmPGH:CvNR:PmG3PDH:OambLPAAT2:PmPGH ::3’Thia4. Proceeding in the 5′ to 3′ direction, bold, lowercase sequences represent genomic DNA from CHK22 that permit targeted integration at the Thia4 locus via homologous recombination, the C. reinhardtii β-tubulin promoter driving expression of the yeast sucrose invertase gene (conferring the ability of CHK22 to metabolize sucrose) is indicated by boxed uppercase text. The initiator ATG and terminator TAA for invertase are indicated by uppercase, bold italics while the coding region is indicated in lowercase italics. The P. moriformis PmPGH 3′-UTR that enables amplification of the OambLPAAT2 gene, is indicated by uppercase underlined text followed by a linker, indicated by lowercase, bold italics, which contains a C. vulgaris nitrate reductase 3′-UTR. The P. moriformis PmG3PDH promoter, indicated by boxed italicized text, drive the expression of the OambLPAAT2. The initiator ATG and terminator TAG codons of the OambLPAAT2 are indicated by uppercase, bold italics, while the remainder of the gene is indicated by bold italics. The PmPGH 3′-UTR is indicated by lowercase underlined text followed by the CHK22 Thia4 genomic region indicated by bold, lowercase text. [0350] Table 109 shows the fatty acid/TAG profiles of seven primary transformants of CHK22 transformed with pCHK1232. The P. moriformis base strain CHK22 is shown as a non-transgenic control. Strains were grown for 120 hrs in 50-mL conical tube at 200 rpm at 28 ^C in lipid production medium. Biomass was harvested, lyophilized to dryness, and subjected to direct transesterification to generate FAMEs for subsequent quantitation and characterization by GC/FID. TAG analysis was conducted using the LCMS method as described above. Table 109: Screen of primary transformants of CHK22 with pCHK1232.
[0351] The amino acid sequences of various LPAATs disclosed herein were compared with the amino acid sequence of CrLPAAT2 to generate a sequence percent identity (%). The results are shown in Table 110. The resulting phenotypes of expressing these LPAATs in the non-transgenic cell line CHK22 is indicated with the OP:OO ratio. [0352] In some embodiments, a microalgal cell provided herein comprises an exogenous gene that encodes for a LPAAT in Table 110. In some embodiments, the exogenous gene can comprise a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a LPAAT in Table 110. Table 110: Percent identity against CrLPAAT2 and OP:OO Ratio
[0353] The amino acid sequences were compared with the amino acid sequence of PedLPAAT2 to generate a sequence percent identity (%). The results are shown in Table 111. The resulted phenotype of expressing these LPAATs in CHK22 is indicated with the OP:OO ratio. [0354] In some embodiments, a microalgal cell provided herein comprises an exogenous gene that encodes for a LPAAT in Table 111. In some embodiments, the exogenous gene can comprise a sequence with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a LPAAT in Table 111. Table 111: Percent identity against PedLPAAT2 and OP:OO Ratio
[0355] Table 112 shows the amino acid sequences of LPAATs described herein. [0356] In some embodiments, a microalgal cell provided herein comprises an exogenous gene that encodes for a LPAAT in Table 112. In some embodiments, the exogenous gene can comprise a sequence that encodes for an enzyme with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a LPAAT in Table 112. In some embodiments, the exogenous gene can comprise a sequence that encodes for an enzyme with at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to any one of SEQ ID NO: 83-106, 155, and 156. Table 112: LPAAT Amino Acid Sequences
[0357] Table 113 shows conserved LPAAT motifs and the percent identity (%) of the most active LPAATs across the entire LPAAT protein relative to PedLPAAT2. The transmembrane domains of these LPAATs were compared and the schematic diagrams of the LPAATs are shown in FIG.11. The LPAAT catalytic domain NHX4D is shown in shaded bar, binding motif EGTR or EGHR is shown in black bar, and transmembrane domains are indicated by gray bar. Table 113: Percent identity against PedLPAAT2 and LPAAT motifs
[0358] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.