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Application of acetyl coenzyme A enhancing module to enhancement of 7-dehydrocholesterol synthesis in microorganisms

A dehydrocholesterol and acetyl coenzyme technology, applied in the field of genetic engineering, can solve the problem of low yield of 7-dehydrocholesterol

Inactive Publication Date: 2015-10-21
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production of 7-dehydrocholesterol in existing recombinant S. cerevisiae strains is low, and further development is needed

Method used

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  • Application of acetyl coenzyme A enhancing module to enhancement of 7-dehydrocholesterol synthesis in microorganisms
  • Application of acetyl coenzyme A enhancing module to enhancement of 7-dehydrocholesterol synthesis in microorganisms
  • Application of acetyl coenzyme A enhancing module to enhancement of 7-dehydrocholesterol synthesis in microorganisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0127] Example 1 The construction of the recombinant Saccharomyces cerevisiae strain producing 7-dehydrocholesterol and the contrast of 7-dehydrocholesterol output

[0128] Source of material:

[0129] pRS425 plasmid: purchased from Addgene (American);

[0130] pRS413 plasmid: purchased from Addgene (American);

[0131] Saccharomyces cerevisiae BY4742 single-gene deletion strain ΔERG5 (MATαhis3Δ1leu2Δ0lys2Δ0ura3Δ0erg5Δ::KanMX): purchased from Thermo Fisher Scientific.

[0132] Acquisition of promoters, terminators and endogenous and exogenous genes

[0133] All promoters (FBA1p, ENO2p, PGI1p, PGK1p, PDC1p), terminators (HXT7t, ADH1t, GPM1t, GPDt, FBA1t, PGK1t) and endogenous genes (alcohol dehydrogenase gene adh2, acetaldehyde dehydrogenase gene ald6, Truncated HMG-CoA reductase gene thmg1): Design appropriate primers with promoters, terminators or gene homologous sequences, and use the Saccharomyces cerevisiae BY4742 genome as a template to amplify by PCR reaction.

[013...

Embodiment 2

[0151] Example 2 Research on the performance of each recombinant strain producing 7-dehydrocholesterol

[0152] The fermentation performances of the recombinant strains SyBE_Sc01100002 and SyBE_Sc01100011 prepared in Example 1 were compared on shake flasks.

[0153] 1. Test material:

[0154] The bacterial strains SyBE_Sc01100002 and SyBE_Sc01100011 prepared in Example 1.

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Abstract

The invention belongs to the technical field of genetic engineering and discloses application of an acetyl coenzyme A enhancing module to enhancement of 7-dehydrocholesterol synthesis in microorganisms. According to the application, the acetyl coenzyme A enhancing module comprises genes expressing ethanol dehydrogenase, genes expressing acetaldehyde dehydrogenase, genes expressing acetyl coenzyme A synthase and genes expressing ATP-citrate lyase. After the acetyl coenzyme A enhancing module is led in the microoranisms, the synthesis amount of the 7-dehydrocholesterol is remarkably increased. A new method is provided for synthesizing 7-dehydrocholesterol, and mass production of 7-dehydrocholesterol is facilitated better.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and particularly relates to the application of an acetyl coenzyme A enhancing module in enhancing the synthesis of 7-dehydrocholesterol in microorganisms. Background technique [0002] Vitamin D is a fat-soluble vitamin and is also regarded as a hormone precursor that acts on calcium and phosphorus metabolism. It is one of the essential vitamins for the human body. According to reports, there are at least ten known types of vitamin D, but the most important is vitamin D 2 and vitamin D 3 . Among them, vitamin D 3 It is the active form of vitamin D with the highest biological metabolism rate. Vitamin D 3 It is mainly synthesized by the body itself: 7-dehydrocholesterol in human skin absorbs UVB in sunlight and can be converted into provitamin D 3 , and then generate a more thermodynamically stable structure through intramolecular rearrangement—vitamin D 3 . Therefore, 7-dehydro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12P33/02C12N15/53C12N15/60C12N15/52C12N1/15
Inventor 肖文海苏皖王颖周晓刘夺元英进
Owner TIANJIN UNIV
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