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Saccharomyces cerevisiae strain with excellent autolysis resistance

A technology of Saccharomyces cerevisiae and microbial strains, applied in the fields of molecular biology and microbiology, can solve the problems of incomplete understanding of yeast autolysis mechanism, and achieve the effect of yeast autolysis

Active Publication Date: 2015-04-22
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the limitations of technology and conditions, people do not have a thorough understanding of the mechanism of yeast autolysis, making these current solutions often unable to fundamentally solve the problem

Method used

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  • Saccharomyces cerevisiae strain with excellent autolysis resistance
  • Saccharomyces cerevisiae strain with excellent autolysis resistance
  • Saccharomyces cerevisiae strain with excellent autolysis resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Construction of embodiment 1 fks1 gene overexpression plasmid

[0026] Saccharomyces cerevisiae model strain W303 (type strain) was used as the host strain, its genome was extracted, the required fragments were amplified by PCR: 18SrDNA-up, 18SrDNA-down, PGK1 and fks1, and the KanMX fragment was amplified using the pUG6 plasmid as a template. The obtained fragments were respectively connected to pMD-19T vector by subcloning, and finally the target fragments were obtained by double enzyme digestion. The whole process of molecular manipulation is as figure 1 shown.

Embodiment 2

[0027] Construction and verification of embodiment 2 recombinant yeast

[0028] The target fragment was introduced into yeast cells by the chemical transformation method in acetic acid, integrated into the yeast genome through homologous recombination, and the transformants were screened by means of G418-resistant replica plates. The result is as figure 2 shown. Extract the transformant genome, and design primers 1 and 2 to screen and verify positive transformants. Primer 1 intercepts the middle part of 18SrDNA as an upstream primer, and the middle part of PGK1 as a downstream primer. If the amplification is successful, there will be 1270bp; primer 2 intercepts KanMX and 18SrDNA The middle position is used as upstream and downstream primers, if successful, there will be a 975bp band.

[0029] The obtained positive transformant was deposited in the General Microorganism Center of China Microbiological Culture Collection Management Committee on September 1, 2014. The preserva...

Embodiment 3

[0030] Embodiment 3 recombinant strain TEM analysis

[0031] In order to visually observe the cell wall changes of the strains, the sections of recombinant bacteria and wild bacteria cells were observed under transmission electron microscope. image 3 It showed that the cell wall of the recombinant bacteria was significantly thicker than that of the wild bacteria. According to the calculation, the cell wall thickness of the wild bacteria is about 200nm, and that of the recombinant bacteria CGMCC No.9628 is about 500nm, twice as much as that of the wild bacteria. The increase of cell wall thickness is due to the accumulation of 1,3-β-glucan content, and the content of 1,3-β-glucan in the cell wall of the recombinant strain CGMCC No.9628 is 62.51% higher than that of the wild strain, which proves that Good effect of gene overexpression method.

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Abstract

The invention discloses a saccharomyces cerevisiae strain with excellent autolysis resistance, and belongs to the field of molecular biology and microbiology. The strain is preserved in China General Microbiological Culture Collection Center (CGMCC) on September 1, 2014, the Preservation No. is CGMCC No. 9628, and the preservation address is Institute of Microbiology at the Chinese Academy of Sciences, at No.3, Courtyard No.1, Beichen West Road, Chaoyang District, Beijing. Compared with original strains, the thickness of cell walls of the strain is doubled, the strain can tolerate a starvation culture for 24 h in a mixture of 50 ng / mL of micafungin, 8% of ethanol, 0.4 M NaCl and sterile water, and autolysis simulation experiment data and scanning electron microscope results (60 h) show that the autolysis of recombinant strains is superior to that of original strains. The strain disclosed by the invention provides a new thought for the better control and utilization of an autolysis phenomenon of saccharomyces cerevisiae, and the obtained strain has a good application value in the aspects of improving the properties such as flavor, foam stability and the like of beer.

Description

technical field [0001] The invention relates to a strain of Saccharomyces cerevisiae strain with excellent anti-autolysis performance, belonging to the fields of molecular biology and microbiology. Background technique [0002] Yeast is one of the most important industrial microorganisms, especially in the field of beer production, known as the soul of beer production. However, in recent years, in the process of beer production, yeast autolysis accompanied by natural aging has seriously affected the quality of beer, such as flavor stability and foam stability. A burning problem. [0003] At present, most of the research on preventing yeast autolysis focuses on improving the beer production process, and a few are improved by optimizing strains. Due to the limitations of technology and conditions, people do not have a thorough understanding of the mechanism of yeast autolysis, making these current solutions often unable to fundamentally solve the problem. [0004] The inven...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81C12C11/00C12R1/865
CPCC12C11/00C12C2200/05C12N9/1051C12N15/81C12Y204/01034
Inventor 李崎李佳王金晶李永仙朱林江刘春凤郑飞云
Owner JIANGNAN UNIV
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