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High-tolerance yeast strain suitable for fermentation of frozen dough and application of high-tolerance yeast strain

A technology of yeast strain and baker's yeast, which is applied in the field of bioengineering, can solve problems such as no relevant reports on freezing tolerance, and achieve the effects of improving the level of production technology, wide application prospects, and promoting popularization and application.

Active Publication Date: 2019-10-22
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the indirect related genetic modification of freezing stress regulation or the unknown regulatory network of freezing stress response. There are no related reports

Method used

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  • High-tolerance yeast strain suitable for fermentation of frozen dough and application of high-tolerance yeast strain
  • High-tolerance yeast strain suitable for fermentation of frozen dough and application of high-tolerance yeast strain
  • High-tolerance yeast strain suitable for fermentation of frozen dough and application of high-tolerance yeast strain

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Experimental program
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Effect test

Embodiment 1

[0033] Example 1: Construction of freeze-tolerant baker's yeast strains

[0034] (1) Construction of expression plasmid Yep-KPS

[0035] The construction process of the expression plasmid Yep-KPS is as follows: figure 1 shown;

[0036] Using the total DNA of yeast strain CICC31616 as a template, the complete sequence of SNF1 gene was amplified by PCR. The reaction system is shown in Table 1:

[0037] Upstream primer 1: 5'-ggaattccagatctcctcgagTGAGCAGTAACAACAACACAAACA-3' (SEQ ID NO: 2);

[0038] Downstream primer 2: 5'-atctatcgcagatccctcgagTCAATTGCTTTGACTGTTAACGG-3' (SEQ ID NO: 3);

[0039] Table 1 PCR amplification of SNF1 gene

[0040]

[0041] The PCR product was connected to the Yep352 plasmid (referred to as Yep-p) containing a strong promoter PGK1 by ClonExpress technology (ClonExpressⅡOne Step Cloning Kit, Vazyme, C211-01) to obtain the vector plasmid Yep-PS with the SNF1 gene:

[0042] The plasmid pUG6 was used as a template to amplify the KanMX gene by PCR. The...

Embodiment 2

[0056] Embodiment 2: Freezing tolerance experiment of recombinant strain and starting strain

[0057] (1) Freezing tolerance test of recombinant strains and starting strains

[0058] Pick a ring of yeast cells in YEPD medium, 30 ℃, static culture for 24h; transfer to molasses medium with 10% (v / v) inoculation amount (add 0.5g / L Ammonium sulfate, 5g / L yeast powder), 30°C, 180r / min, cultivated for 24h to the stationary phase (OD 600 1.5 or so); statically cultivated for 2 hours, centrifuged at 4000r / min for 5 minutes, washed twice with sterile water, and collected the thalli for subsequent use.

[0059] The yeast that above-mentioned centrifugation collects is received respectively by the inoculum size of 2.0% (m / v) simulated dough culture medium (40g / L glucose, 2.5g / L ammonium sulfate, 5g / L urea, 16g / L diphosphate Potassium hydrogen, 5g / L disodium hydrogen phosphate, 0.6g / L magnesium sulfate, 22.5mg / L niacin, 5.0mg / L pantothenic acid, 2.5mg / L vitamin B1, 1.25mg / L vitamin B6, ...

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Abstract

The invention belongs to the technical field of bioengineering, and relates to breeding of industrial microorganisms, in particular to a high-freezing-tolerance accharomyces cerevisiae strain and a construction method thereof. The freezing-tolerance yeast strain is obtained in the mode that in a parent yeast strain, a strong promoter PGK1 is selected for overexpressing a complete sequence of an encoding gene SNF1 of Snf1 protein kinase alpha subunit. Compared with the parent strain, according to the freezing-tolerance accharomyces cerevisiae strain, in a freezing environment, the cell survivalrate and the fermentation capacity of frozen dough are significantly improved, technical obstacles and quality defects in the bread making process are overcome, and the high-freezing-tolerance accharomyces cerevisiae strain has a wide application prospect.

Description

Technical field: [0001] The invention belongs to the technical field of bioengineering and relates to the breeding of industrial microorganisms, in particular to a strain of baker's yeast with high freezing tolerance and a construction method thereof. Background technique: [0002] As a microbial leavening agent and biological loosening agent, baker's yeast is one of the indispensable and important raw materials in the bread production process. Baker's yeast can use the nutrients in the dough to ferment to produce CO 2 , so that the volume of the dough expands and becomes loose; the alcohols and esters produced by fermentation make the bread have a unique flavor and fragrance. my country has a large population, of which about half of the population is mainly pasta, and bread is also diversified. In the development, it is loved by more people. With the development of pasta food, frozen dough technology has been widely promoted, realizing the separation of the pre-production ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81A21D8/04C12R1/865
CPCC12N9/1205C12N15/81A21D8/047
Inventor 林雪刘四新孟露杨旭赵俊珂
Owner HAINAN UNIVERSITY
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