A kind of method for improving Saccharomyces cerevisiae palmitoleic acid content

A technology of Saccharomyces cerevisiae and palmitoleic acid, applied in the biological field, can solve the problems that the expression of fatty acid dehydrogenase cannot be activated, and the regulation effect is not yet clear.

Active Publication Date: 2022-07-26
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mga2 senses the level of unsaturated fatty acids on the cell membrane and binds ligands through the tryptophan residues of the TM transmembrane domain, thereby regulating the transcriptional activity of Mga2, and the higher level of unsaturated fatty acids on the membrane leads to the transcriptional activation of Mga2 Inhibited so that the expression of fatty acid dehydrogenase cannot be activated
However, the regulatory role of its IPT domain and ankyrin repeat sequence on the synthesis of unsaturated fatty acids has not yet been clarified.

Method used

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  • A kind of method for improving Saccharomyces cerevisiae palmitoleic acid content
  • A kind of method for improving Saccharomyces cerevisiae palmitoleic acid content
  • A kind of method for improving Saccharomyces cerevisiae palmitoleic acid content

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1. Effect of overexpression of transcription factor Mga2 on palmitoleic acid content in Saccharomyces cerevisiae

[0057] 1. Construction of genetically engineered bacteria overexpressing transcription factor Mga2

[0058] 1. Construction of Mga2 overexpression vector

[0059] The yeast genome was extracted using Bomed's standard yeast extraction kit to clone the Mga2 gene, and two restriction sites, BamHI and SmaI, were selected and connected to the vector YEplac112 with the strong promoter PGK promoter and CYC terminator of Saccharomyces cerevisiae. -P PGK -T CYC (Shanghai Zeye Biotechnology Co., Ltd., ZY1369).

[0060] The successfully ligated plasmid was transferred into Trans10 competent cells for verification screening and plasmid amplification. Pick a single colony grown on the plate for colony PCR verification, select a single colony that has been successfully verified by colony PCR, and insert it into an LB test tube with Amp resistance for about 12...

Embodiment 2

[0095] Example 2. Effects of overexpression of Mga2 transcription factor and heterologous expression of fatty acid elongase genes KCS and FAE on palmitoleic acid content in Saccharomyces cerevisiae

[0096] 1. Construction of genetically engineered bacteria overexpressing Mga2 transcription factor and heterologously expressing fatty acid elongase gene KCS and FAE

[0097] The fatty acid elongase genes KCS and FAE are from foreign plants, Camelina officinalis and Crabgrass, which are not easy to obtain. Therefore, the sequences were sent to Suzhou Jinweizhi Company for direct synthesis and linked to YEplac112-P PGK -T CYC on the carrier.

[0098] The nucleotide sequence of the fatty acid elongase gene KCS is sequence 2 in the sequence listing;

[0099] The nucleotide sequence of the fatty acid elongase gene FAE is sequence 3 in the sequence listing.

[0100] 1. Construction of MGA2 and KCS double gene expression engineering bacteria YS07

[0101] pYES2-P TEF1 -MGA2 706aa ...

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Abstract

The invention discloses a method for increasing the palmitoleic acid content of Saccharomyces cerevisiae. The present invention provides recombinant bacteria, in order to carry out the following 1) transformation in the oil-producing Saccharomyces cerevisiae to obtain the recombinant bacteria: 1) the recombinant bacteria obtained by improving the transcriptional activity of the OLE1 gene in the oil-producing Saccharomyces cerevisiae. In the present invention, in the Saccharomyces cerevisiae strain YS58 that produces more oil, the transcription factor Mga2 or the truncated Mga2 gene fragment is overexpressed, the expression level of the OLE1 gene regulating Δ9 desaturase is increased, and the content of palmitoleic acid in Saccharomyces cerevisiae is increased, Simultaneously express the exogenous fatty acid elongase genes KCS and FAE to construct Saccharomyces cerevisiae genetically engineered bacteria and increase the content of palmitoleic acid in Saccharomyces cerevisiae.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for increasing the palmitoleic acid content of Saccharomyces cerevisiae. Background technique [0002] Palmitoleic acid is a 16-carbon monounsaturated fatty acid with a double bond at the 7th carbon atom at the carbon end (C16:1n-7). It has a wide range of pharmaceutical and cosmetic applications, has certain curative effects on chronic diseases such as obesity, diabetes, and fatty liver, and has application potential in hypoglycemic, anti-inflammatory, skin pigmentation improvement and food fields. At present, seafood such as fish oil is the main source of palm oleic acid, but due to the international ban on whaling and the lack of fishery resources, the source of palm oleic acid is limited. Efforts have therefore been made to expand the possibility of isolating this fatty acid from microbial sources. [0003] As a biological metabolite, palmitoleic acid mainly exists in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/19C12N9/02C12N9/10C12P7/6409C12R1/865
CPCC12N9/0071C12N9/1029C12P7/6409C12Y114/19001C12Y203/01086
Inventor 刘军锋庞杰邓利王芳
Owner BEIJING UNIV OF CHEM TECH
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