Title:
PRODUCTION OF TYRIAN PURPLE CELL DYE BY USING ESCHERICHIA COLI AND DYEING METHOD USING SAME
Document Type and Number:
WIPO Patent Application WO/2022/097856
Kind Code:
A1
Abstract:
Tyrian purple, mainly consisting of 6,6'-dibromoindigo (6BrIG), is an ancient dye extracted from sea snails and has recently been demonstrated as a biocompatible semiconductor material. However, the synthesis of Tyrian purple is limited due to uncharacterized biosynthetic pathways and the difficulty of regiospecific bromination. The present invention presents an effective 6BrIG production strategy in Escherichia coli using tryptophan 6-halogenase (SttH), tryptophanase (TnaA), and flavin-containing monooxygenase (MaFMO). Since tryptophan halogenases are expressed in highly insoluble forms in Escherichia coli, a flavin reductase Fre that regenerates FADH2 for the halogenase reaction was used as an N-terminal soluble tag of SttH. A consecutive two-cell reaction system of the present invention is designed to enable the mass production of regiospecifically brominated precursors of 6BrIG by spatiotemporal separation of bromination and bromotryptophan degradation. Through these approaches, in the present invention, 315.0 mg/L of 6BrIG was produced from tryptophan, and the successful synthesis of regiospecifically dihalogenated indigos was induced. In addition, the present invention shows that 6BrIG-containing Escherichia coli cells can be directly used as a cell dye.
Inventors:
LEE JEONG CHAN (KR)
KIM BYUNG GEE (KR)
KIM JOON WON (KR)
KIM BYUNG GEE (KR)
KIM JOON WON (KR)
Application Number:
PCT/KR2021/005504
Publication Date:
May 12, 2022
Filing Date:
April 30, 2021
Export Citation:
Assignee:
SEOUL NAT UNIV R&DB FOUNDATION (KR)
International Classes:
C12N15/70; C09B7/00; C12N9/02; D06P1/22
Domestic Patent References:
| WO2020015839A1 | 2020-01-23 |
Foreign References:
| US4752301A | 1988-06-21 |
Other References:
LEE JEONGCHAN; KIM JOONWON; SONG JI EUN; SONG WON-SUK; KIM EUN-JUNG; KIM YUN-GON; JEONG HEE-JIN; KIM HYE RIM; CHOI KWON-YOUNG; KIM: "Production of Tyrian purple indigoid dye from tryptophan in", NATURE CHEMICAL BIOLOGY, NATURE PUBLISHING GROUP US, NEW YORK, vol. 17, no. 1, 1 January 1900 (1900-01-01), New York, pages 104 - 112, XP037317216, ISSN: 1552-4450, DOI: 10.1038/s41589-020-00684-4
JIA ZENG; JIXUN ZHAN: "Characterization of a tryptophan 6-halogenase from", BIOTECHNOLOGY LETTERS, SPRINGER NETHERLANDS, DORDRECHT, vol. 33, no. 8, 19 March 2011 (2011-03-19), Dordrecht , pages 1607 - 1613, XP019926090, ISSN: 1573-6776, DOI: 10.1007/s10529-011-0595-7
FABARA ANDREA N.; FRAAIJE MARCO W.: "Production of indigo through the use of a dual-function substrate and a bifunctional fusion enzyme", ENZYME AND MICROBIAL TECHNOLOGY, STONEHAM, MA, US, vol. 142, 14 October 2020 (2020-10-14), US , XP086353038, ISSN: 0141-0229, DOI: 10.1016/j.enzmictec.2020.109692
LEE JEONGCHAN; KIM JOONWON; SONG JI EUN; SONG WON-SUK; KIM EUN-JUNG; KIM YUN-GON; JEONG HEE-JIN; KIM HYE RIM; CHOI KWON-YOUNG; KIM: "Production of Tyrian purple indigoid dye from tryptophan in", NATURE CHEMICAL BIOLOGY, NATURE PUBLISHING GROUP US, NEW YORK, vol. 17, no. 1, 1 January 1900 (1900-01-01), New York, pages 104 - 112, XP037317216, ISSN: 1552-4450, DOI: 10.1038/s41589-020-00684-4
KIM, J.Y ET AL.: "In vitro characterization of CYP102G4 from Streptomyces cattleya: A self-sufficient P450 naturally producing indigo", BIOCHIM BIOPHYS ACTA PROTEINS PROTEOM, vol. 1866, 2018, pages 60 - 67
ZHANG, X.W ET AL.: "Cloning and expression of naphthalene dioxygenase genes from Comamonas sp MQ for indigoids production", PROCESS BIOCHEM, vol. 48, 2013, pages 581 - 587, XP028546630, DOI: 10.1016/j.procbio.2013.02.008
KIM, J.Y ET AL.: "Production of dyestuffs from indole derivatives by naphthalene dioxygenase and toluene dioxygenase", LETT APPL MICROBIOL, vol. 36, 2003, pages 343 - 348
NAMGUNG, S ET AL.: "Ecofriendly one-pot biosynthesis of indigo derivative dyes using CYP102G4 and PrnA halogenase", DYES PIGMENTS, vol. 162, 2019, pages 80 - 88
HAN, G.H ET AL., ENZYME MICROB TECH, vol. 42, 2008, pages 617 - 623
KIM, H.J. ET AL., DYES PIGMENTS, vol. 140, 2017, pages 29 - 35
JIA ZENG; JIXUN ZHAN: "Characterization of a tryptophan 6-halogenase from", BIOTECHNOLOGY LETTERS, SPRINGER NETHERLANDS, DORDRECHT, vol. 33, no. 8, 19 March 2011 (2011-03-19), Dordrecht , pages 1607 - 1613, XP019926090, ISSN: 1573-6776, DOI: 10.1007/s10529-011-0595-7
FABARA ANDREA N.; FRAAIJE MARCO W.: "Production of indigo through the use of a dual-function substrate and a bifunctional fusion enzyme", ENZYME AND MICROBIAL TECHNOLOGY, STONEHAM, MA, US, vol. 142, 14 October 2020 (2020-10-14), US , XP086353038, ISSN: 0141-0229, DOI: 10.1016/j.enzmictec.2020.109692
LEE JEONGCHAN; KIM JOONWON; SONG JI EUN; SONG WON-SUK; KIM EUN-JUNG; KIM YUN-GON; JEONG HEE-JIN; KIM HYE RIM; CHOI KWON-YOUNG; KIM: "Production of Tyrian purple indigoid dye from tryptophan in", NATURE CHEMICAL BIOLOGY, NATURE PUBLISHING GROUP US, NEW YORK, vol. 17, no. 1, 1 January 1900 (1900-01-01), New York, pages 104 - 112, XP037317216, ISSN: 1552-4450, DOI: 10.1038/s41589-020-00684-4
KIM, J.Y ET AL.: "In vitro characterization of CYP102G4 from Streptomyces cattleya: A self-sufficient P450 naturally producing indigo", BIOCHIM BIOPHYS ACTA PROTEINS PROTEOM, vol. 1866, 2018, pages 60 - 67
ZHANG, X.W ET AL.: "Cloning and expression of naphthalene dioxygenase genes from Comamonas sp MQ for indigoids production", PROCESS BIOCHEM, vol. 48, 2013, pages 581 - 587, XP028546630, DOI: 10.1016/j.procbio.2013.02.008
KIM, J.Y ET AL.: "Production of dyestuffs from indole derivatives by naphthalene dioxygenase and toluene dioxygenase", LETT APPL MICROBIOL, vol. 36, 2003, pages 343 - 348
NAMGUNG, S ET AL.: "Ecofriendly one-pot biosynthesis of indigo derivative dyes using CYP102G4 and PrnA halogenase", DYES PIGMENTS, vol. 162, 2019, pages 80 - 88
HAN, G.H ET AL., ENZYME MICROB TECH, vol. 42, 2008, pages 617 - 623
KIM, H.J. ET AL., DYES PIGMENTS, vol. 140, 2017, pages 29 - 35
Attorney, Agent or Firm:
KOREANA PATENT FIRM (KR)
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