Methods and materials for biosynthesis of manoyl oxide
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12P-017/06
C12N-009/88
C12P-005/00
C12P-017/04
C07D-307/92
C07D-311/92
C12N-009/10
출원번호
US-0523868
(2015-11-13)
등록번호
US-10208326
(2019-02-19)
우선권정보
DK-2014 70694 (2014-11-13)
국제출원번호
PCT/EP2015/076595
(2015-11-13)
국제공개번호
WO2016/075302
(2016-05-19)
발명자
/ 주소
Jensen, Niels Bjerg
출원인 / 주소
EVOLVA SA
대리인 / 주소
McDonnell Boehnen Hulbert & Berghoff LLP
인용정보
피인용 횟수 :
0인용 특허 :
4
초록
The invention relates to recombinant microorganisms and methods for producing manoyl oxide.
대표청구항▼
1. A method of producing a manoyl oxide, comprising: growing a recombinant host, comprising: (a) a gene encoding a geranylgeranyl diphosphate synthase (GGPPS) polypeptide, wherein the GGPPS polypeptide comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth
1. A method of producing a manoyl oxide, comprising: growing a recombinant host, comprising: (a) a gene encoding a geranylgeranyl diphosphate synthase (GGPPS) polypeptide, wherein the GGPPS polypeptide comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NOs:2 or 4;(b) a gene encoding a polypeptide catalyzing formation of copal-8-ol diphosphate from qeranylgeranyl diphosphate (GGPP), wherein the polypeptide catalyzing formation of copal-8-ol diphosphate from GGPP comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:6; and(c) a gene encoding a polypeptide catalyzing formation of manoyl oxide from copal-8-ol diphosphate, wherein the polypeptide catalyzing formation of manoyl oxide from copal-8-ol diphosphate comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NOs:9 or 11 in a culture medium, under conditions in which the genes are expressed;wherein the manoyl oxide is synthesized by the host. 2. The method of claim 1, wherein the manoyl oxide is (13R) manoyl oxide. 3. A method for producing a terpenoid, comprising: (a) culturing a recombinant host capable of producing a manoyl oxide, comprising: (i) a gene encoding a geranylgeranyl diphosphate synthase (GGPPS) polypeptide, wherein the GGPPS polypeptide comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NOs:2 or 4;(ii) a gene encoding a polypeptide catalyzing formation of copal-8-ol diphosphate from geranylgeranyl diphosphate (GGPP), wherein the polypeptide catalyzing formation of copal-8-ol diphosphate from GGPP comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:6; and(iii) a gene encoding a polypeptide catalyzing formation of manoyl oxide from copal-8-ol diphosphate, wherein the polypeptide catalyzing formation of manoyl oxide from copal-8-ol diphosphate comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NOs:9 or 11;wherein at least one of the genes is a recombinant gene;in a culture medium, under conditions in which the genes are expressed; wherein the manoyl oxide is synthesized by the recombinant host;(b) isolating the manoyl oxide produced by the recombinant host; and(c) converting the manoyl oxide into the terpenoid. 4. The method of claim 3, wherein the manoyl oxide is isolated from the recombinant host and/or from the culture medium. 5. The method of claim 3, wherein the manoyl oxide is converted into the terpenoid by organic chemical synthesis. 6. The method of claim 3, wherein the terpenoid is forskolin. 7. The method of claim 3, wherein the terpenoid is ambrox. 8. The method of claim 1, further comprising quantifying the manoyl oxide produced by the recombinant host. 9. The method of claim 1, further comprising isolating the manoyl oxide produced by the recombinant host. 10. The method of claim 3, wherein the recombinant host is a plant cell, a mammalian cell, an insect cell, a fungal cell, or a bacterial cell. 11. The method of claim 10, wherein the bacterial cell is a Escherichia bacteria cell, a Lactobacillus bacteria cell, a Lactococcus bacteria cell, a Cornebacterium bacteria cell, a Acetobacter bacteria cell, a Acinetobacter bacteria cell or a Pseudomonas bacterial cell. 12. The method of claim 10, wherein the fungal cell comprises a yeast cell. 13. The method of claim 12, wherein the yeast cell is a cell from Saccharomyces cerevisiae, Schizosaccharomyces pombe, Yarrowia lipolytica, Candida glabrata, Ashbya gossypii, Cyberlindnera jadinii, Pichia pastoris, Kluyveromyces lactis, Hansenula polymorpha, Candida boidinii, Arxula adeninivorans, Xanthophyllomyces dendrorhous or Candida albicans species. 14. The method of claim 12, wherein the yeast cell is a Saccharomycete. 15. The method of claim 12, wherein the yeast cell is a cell from the Saccharomyces cerevisiae species.
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이 특허에 인용된 특허 (4)
Lundquist Ronald C. ; Walters David A., Fertile transgenic corn plants.
Sanford John C. (Geneva) DeVit Michael J. (Geneva NY) Bruner Ronald F. (Sewell NJ) Johnston Stephen A. (Durham NC), Method and apparatus for introducing biological substances into living cells.
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