Title:
Methods of optimizing substrate pools and biosynthesis of poly-beta-hydrdoxybutyrate-copoly-beta-hydroxyvalerate in bacteria and plants
Document Type and Number:
WIPO Patent Application WO1998000557
Kind Code:
A3
Abstract:
Genes and methods for optimizing levels of substrates employed in the biosynthesis of copolymers of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) in plants and bacteria via manipulation of normal metabolic pathways using recombinant DNA techniques are provided. This is achieved through the use of a variety of wild-type and/or deregulated enzymes involved in the biosynthesis of aspartate family amino acids, and wild-type or deregulated forms of enzymes, such as threonine deaminase, involved in the conversion of threonine to P(3HB-co-3HV) copolymer endproduct. By these methods, enhanced levels of threonine, alpha -ketobutyrate, propionate, propionyl-CoA, beta -ketovaleryl-CoA, and beta -hydroxyvaleryl-CoA are produced. Also provided are methods for the biological production of P(3HB-co-3HV) copolymers in plants and bacteria utilizing propionyl-CoA produced through a variety of engineered metabolic pathways. Introduction into plants and bacteria of an appropriate beta -ketothiolase, beta -ketoacyl-CoA reductase, and PHA synthase, alone or in combination with various enzymes involved in asparate family amino acid biosynthesis and the conversion of threonine to PHA copolymer precursors, will permit these organisms to produce P(3HB-co-3HV) copolymers.
Inventors:
GRUYS KENNETH JAMES (US)
MITSKY TIMOTHY ALBERT (US)
KISHORE GANESH MURTHY (US)
SLATER STEVEN CHARLES (US)
PADGETTE STEPHEN ROGERS (US)
STARK DAVID MARTIN (US)
HINCHEE MAUD ANN WRIGHTSON (US)
CLEMENTE THOMAS ELMO (US)
CONNOR-WARD DANNETTE VAUDRILYN (US)
FEDELE MARY JACQUELINE (US)
FRY JOYCE ELLEN (US)
HOWE ARLENE RENE (US)
ROZMAN RENEE JEAN (US)
MITSKY TIMOTHY ALBERT (US)
KISHORE GANESH MURTHY (US)
SLATER STEVEN CHARLES (US)
PADGETTE STEPHEN ROGERS (US)
STARK DAVID MARTIN (US)
HINCHEE MAUD ANN WRIGHTSON (US)
CLEMENTE THOMAS ELMO (US)
CONNOR-WARD DANNETTE VAUDRILYN (US)
FEDELE MARY JACQUELINE (US)
FRY JOYCE ELLEN (US)
HOWE ARLENE RENE (US)
ROZMAN RENEE JEAN (US)
Application Number:
PCT/US1997/008983
Publication Date:
July 23, 1998
Filing Date:
May 28, 1997
Export Citation:
Assignee:
MONSANTO CO (US)
International Classes:
C12N1/21; C12N9/00; C12N9/04; C12N9/10; C12N9/88; C12N15/54; C12N15/82; C12P7/62; (IPC1-7): C12N15/82; C12N15/54; C12N15/53; C12N15/52; C12N15/60; C12N9/10; C12N5/10; A01H5/00; C12N1/21; C12P7/62
Domestic Patent References:
| WO1993006225A1 | 1993-04-01 | |||
| WO1995005472A2 | 1995-02-23 | |||
| WO1991018995A1 | 1991-12-12 | |||
| WO1995019442A1 | 1995-07-20 | |||
| WO1987002984A1 | 1987-05-21 | |||
| WO1994011519A1 | 1994-05-26 | |||
| WO1992019747A1 | 1992-11-12 |
Foreign References:
| US5416011A | 1995-05-16 |
Other References:
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FRY, J., ET AL.: "Transformation of Brassica napus with Agrobacterium based vectors", PLANT CELL REPORTS, vol. 6, 1987, pages 321 - 325, XP002045226
POIRIER, Y., ET AL.: "Production of polyhydroxyalkanoates, a family of biodegradable plastics and elastomers, in bacteria and plants", BIOTECHNOLOGY, vol. 13, 1995, pages 142 - 150, XP002045222
MOURAD, G., ET AL.: "L-O-methylthreonine-resistant mutant of Arabidopsis defective in isoleucine feedback regulation", PLANT PHYSIOLOGY, vol. 107, 1995, pages 43 - 52, XP002045223
HINCHEE, M.A.W., ET AL.: "Production of transgenic soybean plants using Agrobacterium-mediated DNA transfer.", BIOTECHNOLOGY, vol. 6, August 1988 (1988-08-01), pages 915 - 922, XP002045224
RADKE, S.E., ET AL.: "Transformation of Brassica napus L., using Agrobacterium tumefaciens: developmentally regulated expression of a reintroduced napin gene", THEOR. APPL. GENET., vol. 75, 1988, pages 685 - 694, XP002045225
MARTON L ET AL: "FACILE TRANSFORMATION OF ARABIDOPSIS.", PLANT CELL REP 10 (5). 1991. 235-239., XP002060527
DATABASE DISSABS BUNNAG, SUMONTHIP [PH.D.]: "SOMACLONAL VARIATION, REGENERATION AND TRANSFORMATION OF QUINCE (CYDONIA OBLONGA MILL.) AND PEAR (PYRUS COMMUNIS L.)", XP002060528
DOI Y: "MICROBIAL SYNTHESIS, PHYSICAL PROPERTIES, AND BIODEGRADABILITY OF POLYHYDROXYALKANOATES", MACROMOLECULAR SYMPOSIA, vol. 98, 1 July 1995 (1995-07-01), pages 585 - 599, XP000543439
POIRIER, Y., ET AL.: "Perspectives on the production of polyhydroxyalkanoates in plants", FEMS MICRBIOL. REV., vol. 103, 1992, pages 237 - 246, XP002045227
NAWRATH, C., ET AL.: "Targeting of the polyhydroxybutyrate biosynthetic pathway to the plastids of Arabidopsis thaliana results in high levels of polymer accumulation", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA., vol. 91, December 1994 (1994-12-01), WASHINGTON US, pages 12760 - 12764, XP002045228
EISENSTEIN, E., ET AL.: "An expanded two-state model accounts for homotropic cooperativity in biosynthetic threonine deaminase from Escherichia coli", BIOCHEMISTRY, vol. 34, 1995, pages 9403 - 9412, XP002045220
HAYWOOD, G.W., ET AL.: "Characterisation of two 3-ketothiolases possessing differing substrate specificities in the polyahydroxyalkanoate synthesising organism Alcaligenes eutrophus", FEMS MICRBIOL. LETT., vol. 52, 1988, pages 91 - 96, XP002045221
FRY, J., ET AL.: "Transformation of Brassica napus with Agrobacterium based vectors", PLANT CELL REPORTS, vol. 6, 1987, pages 321 - 325, XP002045226
POIRIER, Y., ET AL.: "Production of polyhydroxyalkanoates, a family of biodegradable plastics and elastomers, in bacteria and plants", BIOTECHNOLOGY, vol. 13, 1995, pages 142 - 150, XP002045222
MOURAD, G., ET AL.: "L-O-methylthreonine-resistant mutant of Arabidopsis defective in isoleucine feedback regulation", PLANT PHYSIOLOGY, vol. 107, 1995, pages 43 - 52, XP002045223
HINCHEE, M.A.W., ET AL.: "Production of transgenic soybean plants using Agrobacterium-mediated DNA transfer.", BIOTECHNOLOGY, vol. 6, August 1988 (1988-08-01), pages 915 - 922, XP002045224
RADKE, S.E., ET AL.: "Transformation of Brassica napus L., using Agrobacterium tumefaciens: developmentally regulated expression of a reintroduced napin gene", THEOR. APPL. GENET., vol. 75, 1988, pages 685 - 694, XP002045225
MARTON L ET AL: "FACILE TRANSFORMATION OF ARABIDOPSIS.", PLANT CELL REP 10 (5). 1991. 235-239., XP002060527
DATABASE DISSABS BUNNAG, SUMONTHIP [PH.D.]: "SOMACLONAL VARIATION, REGENERATION AND TRANSFORMATION OF QUINCE (CYDONIA OBLONGA MILL.) AND PEAR (PYRUS COMMUNIS L.)", XP002060528
DOI Y: "MICROBIAL SYNTHESIS, PHYSICAL PROPERTIES, AND BIODEGRADABILITY OF POLYHYDROXYALKANOATES", MACROMOLECULAR SYMPOSIA, vol. 98, 1 July 1995 (1995-07-01), pages 585 - 599, XP000543439
POIRIER, Y., ET AL.: "Perspectives on the production of polyhydroxyalkanoates in plants", FEMS MICRBIOL. REV., vol. 103, 1992, pages 237 - 246, XP002045227
NAWRATH, C., ET AL.: "Targeting of the polyhydroxybutyrate biosynthetic pathway to the plastids of Arabidopsis thaliana results in high levels of polymer accumulation", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA., vol. 91, December 1994 (1994-12-01), WASHINGTON US, pages 12760 - 12764, XP002045228
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