Method for producing 3-hydroxypropionic acid and other products
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12P-007/62
A61L-015/24
C12M-001/00
C12N-009/02
C12P-007/40
C12P-007/42
C12P-007/52
C12N-015/63
출원번호
US-0575927
(2014-12-18)
등록번호
US-9428778
(2016-08-30)
발명자
/ 주소
Lynch, Michael D.
Gill, Ryan T.
Lipscomb, Tanya E.W.
출원인 / 주소
Cargill, Incorporated
인용정보
피인용 횟수 :
5인용 특허 :
69
초록
This invention relates to metabolically engineered microorganism strains, such as bacterial strains, in which there is an increased utilization of malonyl-CoA for production of a chemical product, which includes 3-hydroxypropionic acid.
대표청구항▼
1. A method for producing a product having malonyl-CoA as a substrate in a microbial production pathway of the product, the method comprising combining a carbon source and a microorganism cell culture to produce said product, wherein a) said cell culture comprises an inhibitor of a fatty acid syntha
1. A method for producing a product having malonyl-CoA as a substrate in a microbial production pathway of the product, the method comprising combining a carbon source and a microorganism cell culture to produce said product, wherein a) said cell culture comprises an inhibitor of a fatty acid synthase or said microorganism is genetically modified for reduced enzymatic activity of an enzyme wherein the enzyme is selected from the group consisting of beta-ketoacyl-ACP synthase (fabH, fabF, fabB), beta-ketoacyl-ACP reductase (fabG), beta-hydroxyacyl-ACP dehydratase (fab A), and enoyl-ACP reductase (fabI); and either:b1) said microorganism is genetically modified for overexpression of udhA or pntAB; orb2) said microorganism is genetically modified for overexpression of accA, accB, accC, accD, or any combination thereof. 2. The method of claim 1, wherein said carbon source has a ratio of carbon-14 to carbon-12 of about 1.0×10−14 or greater. 3. The method of claim 1, wherein said carbon source is glucose, sucrose, fructose, dextrose, lactose, or a combination thereof. 4. The method of claim 1, wherein said inhibitor of the fatty acid synthase is selected from the group consisting of thiolactomycin, triclosan, cerulenin, thienodiazaborine, and isoniazid. 5. The method of claim 1, wherein said reduced enzymatic activity of the enzyme occurs via introduction of a heterologous nucleic acid sequence comprising an inducible promoter operably linked to a sequence coding for the enzyme or a heterologous nucleic acid sequence coding for the enzyme with reduced activity. 6. The method of claim 5, wherein said beta-ketoacyl-ACP synthase has temperature sensitive enzyme activity or said enoyl-ACP reductase has temperature sensitive enzyme activity. 7. The method of claim 1, wherein said overexpression of udhA or pntAB occurs by introduction of a heterologous nucleic acid sequence coding for a polypeptide having at least 90% homology with a sequence selected from the group consisting of SEQ ID NO. 776 and SEQ ID NO. 778. 8. The method of claim 1, wherein said microorganism is further genetically modified for increased intracellular bicarbonate levels by introduction of a heterologous nucleic acid sequence coding for a polypeptide having cyanase and/or carbonic anhydrase activity. 9. The method of claim 8, wherein said heterologous nucleic acid sequence is selected from a sequence having at least 90% homology with SEQ ID NO. 337. 10. The method of claim 1, wherein said overexpression of accA, accB, accC, accD, or any combination thereof occurs by introduction of a heterologous nucleic acid sequence coding for a polypeptide having at least 90% homology with a sequence selected from the group consisting of SEQ ID NO. 768, SEQ ID NO.769, SEQ ID NO.770, SEQ ID NO.771, SEQ ID NO.772, SEQ ID NO.773, SEQ ID NO.774, and SEQ ID NO. 775. 11. The method of claim 1, wherein said microorganism is further genetically modified to decrease activity of lactate dehydrogenase, phosphate acetyltransferase, pyruvate oxidase, pyruvate-formate lyase, or a combination thereof. 12. The method of claim 1, wherein said microorganism cell culture comprises increased intracellular bicarbonate levels by supplementation with bicarbonate or carbonate. 13. The method of claim 1, wherein the product comprises an alkyl carboxylate. 14. The method of claim 13, wherein the alkyl carboxylate is selected from the group consisting of: a methyl ester of a carboxylic acid, an ethyl ester of a carboxylic acid, and mixtures thereof. 15. The method of claim 14, wherein the methyl ester of a carboxylic acid comprises a methyl ester of 3-hydroxypropionic acid and the ethyl ester of a carboxylic acid comprises an ethyl ester of 3-hydroxypropionic acid. 16. The method of claim 13, wherein the alkyl carboxylate is selected from the group consisting of methyl carboxylates and ethyl carboxylates, and mixtures thereof. 17. The method of claim 16, wherein the methyl carboxylate is a methyl ester of 3-hydroxypropionic acid and the ethyl carboxylate is an ethyl ester of 3-hyrdroxypropionic acid.
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