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Method for increasing content of palmitoleic acid in saccharomyces cerevisiae

A technology of Saccharomyces cerevisiae and palmitoleic acid, applied in the biological field, can solve the problems such as the unclear regulation function and the inability to activate the expression of fatty acid dehydrogenase

Active Publication Date: 2021-06-08
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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  • Method for increasing content of palmitoleic acid in saccharomyces cerevisiae
  • Method for increasing content of palmitoleic acid in saccharomyces cerevisiae
  • Method for increasing content of palmitoleic acid in saccharomyces cerevisiae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1, the effect of overexpression of transcription factor Mga2 on palmitoleic acid content of Saccharomyces cerevisiae

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

[0058] 1. Construction of Mga2 overexpression vector

[0059] Use the standard yeast extraction kit of Bomed Company to extract the yeast genome to clone the Mga2 gene, select two restriction sites of BamHI and SmaI and connect it to the vector YEplac112 with a strong promoter of Saccharomyces cerevisiae PGK promoter and a CYC terminator -P PGK -T CYC (Shanghai Zeye Biotechnology Co., Ltd., ZY1369).

[0060] The successfully ligated plasmids were transferred into Trans10 competent cells for verification screening and amplification of the plasmids. Pick a single colony grown on the plate for colony PCR verification, select a single colony with successful PCR verification and insert it into an LB test tube with Amp resistance for about 12 hours,...

Embodiment 2

[0095] Example 2. Effect of Overexpression of Mga2 Transcription Factor and Simultaneous Expression of Fatty Acid Elongase Genes KCS and FAE on Palmitoleic Acid Content of Saccharomyces cerevisiae

[0096] 1. Construction of overexpressed Mga2 transcription factor and simultaneous heterologous expression of fatty acid elongase gene KCS and FAE genetically engineered bacteria

[0097] Fatty acid elongase genes KCS and FAE are from the foreign plants Michelina and Crabapple and are not easy to obtain. Therefore, the sequences were sent to Suzhou Jinweizhi Co., Ltd. for direct synthesis and ligated in 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 list.

[0100] 1. Construction of MGA2 and KCS dual-gene expression engineering strain YS07

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

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Abstract

The invention discloses a method for increasing the content of palmitoleic acid in saccharomyces cerevisiae. The invention provides a recombinant bacterium which is obtained by performing the following transformation in the oil-producing saccharomyces cerevisiae: 1) the recombinant bacterium is obtained by improving the transcriptional activity of an OLE1 gene in the oil-producing saccharomyces cerevisiae. In a saccharomyces cerevisiae strain YS58 which produces more oil, a transcription factor Mga2 or a truncated Mga2 gene segment is overexpressed, and the expression level of an OLE1 gene for regulating delta 9 desaturase is improved, so that the content of the palmitoleic acid in the saccharomyces cerevisiae is improved, exogenous fatty acid elongase genes KCS and FAE are expressed at the same time, a saccharomyces cerevisiae genetically engineered bacterium is constructed, and the content of the palmitoleic acid in the saccharomyces cerevisiae is increased.

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 seventh carbon atom at the carbon end (C16:1n-7). It has a wide range of medical and cosmetic applications, and has certain curative effects on chronic diseases such as obesity, diabetes, and fatty liver. It has application potential in the fields of lowering blood sugar, anti-inflammation, improving skin pigmentation, and food. At present, seafood such as fish oil is the main source of palmitoleic acid, but due to the international ban on whaling and the lack of fishery resources, the source of palmitoleic acid is limited. Efforts have therefore been made to expand the possibilities for isolating such fatty acids from microbial sources. [0003] As a biological metabolite, palmitoleic ac...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12N9/02C12N9/10C12P7/64C12R1/865
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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