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A high tolerance yeast strain suitable for frozen dough fermentation and its application

A technology of yeast strain and frozen dough, 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, promoting popularization and application, and widening application prospects.

Active Publication Date: 2022-02-22
HAINAN UNIV
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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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  • A high tolerance yeast strain suitable for frozen dough fermentation and its application
  • A high tolerance yeast strain suitable for frozen dough fermentation and its application
  • A high tolerance yeast strain suitable for frozen dough fermentation and its application

Examples

Experimental program
Comparison scheme
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 the breeding of industrial microorganisms, in particular to a strain of baker's yeast with high freezing tolerance and a construction method thereof. The freeze-tolerant yeast strain is obtained by selecting a strong promoter PGK1 to overexpress the entire sequence of the coding gene SNF1 of the alpha subunit of Snf1 protein kinase in the parental yeast strain. Compared with the parent strain, the cell survival rate of the strain and the fermentability of frozen dough in the frozen environment are significantly improved, which solves the technical obstacles and quality defects in the bread making process, and 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 Patents(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 UNIV
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