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Freezing-resistant yeast strain for bread fermentation and breeding method thereof

A yeast strain, freeze-resistant technology, applied in the field of bioengineering, can solve the problems of restricting the development of frozen dough technology, rough structure, insufficient dough expansion, etc., and achieve the effect of improving the synthesis ability of trehalose, wide application prospects, and not easy to recover mutations

Active Publication Date: 2014-01-08
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

After being frozen for 20 days, the vitality of ordinary baker’s yeast has dropped by more than half, resulting in insufficient expansion of the thawed dough, prolonging the proofing time, and the finished bread is small in size, rough in structure, poor in taste, and the quality of the product has dropped significantly
It is the existence of these defects that limits the development of frozen dough technology
In my country, there are many researches on the technology of frozen dough, but there are few researches on the mechanism of baker's yeast freezing resistance and strain selection, especially in the aspect of freezing resistant yeast products, which not only restricts the development of yeast industry, but also It also restricts the popularization and application of frozen dough technology in my country

Method used

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  • Freezing-resistant yeast strain for bread fermentation and breeding method thereof
  • Freezing-resistant yeast strain for bread fermentation and breeding method thereof
  • Freezing-resistant yeast strain for bread fermentation and breeding method thereof

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Embodiment 1

[0022] Embodiment 1: Breeding of freeze-resistant yeast strain

[0023] (1) Construction of recombinant strains

[0024] The construction process of recombinant strains is as follows: figure 1 shown. The diploid starting strain CICC31616 was haploidized to generate a-type and α-type haploid strains 70a and 17α, and the recombinant haploid strains were constructed by two homologous recombination methods, and the KanMX resistance gene was removed and the haploid strains were hybridized Somatic strains to generate diploid recombinant strain BT-N.

[0025] The construction process of the recombinant plasmid pUC-TNBAK is as follows: figure 2 and image 3 shown. By PCR amplification technology, the sequence NA and NB fragments on both sides of the NTH1 gene for homologous recombination to knock out the gene and the KanMX resistance gene from the pUG6 plasmid were amplified, and the three fragments were amplified according to the pUC19 plasmid as the vector The sequence ligati...

Embodiment 2

[0037] Embodiment 2: Fermentation experiment of freezing-resistant yeast strain

[0038] (1) Freezing tolerance experiments of transformants and haploid parents

[0039]The haploid parents 70a and 17α and their corresponding recombinant haploids were simultaneously subjected to liquid simulated dough medium freezing experiments. After freezing at -20°C for 21 days, the cell survival rate and relative fermentation ability were measured. The results are shown in Table 1. The results showed that the intracellular trehalose content of the parent 70a was 95mg / g, the freezing survival rate was 25%, and the relative fermentability was 23.22%. The intracellular trehalose content of the recombinant haploid was 118mg / g, and the freezing survival rate was 83.78%, the relative fermentability is 47.06%, that is, the intracellular trehalose content of the recombinant haploid ΔN+TPS1a increased by 23mg / g compared with the parent 70a, the cell freezing survival rate increased by 58.78%, and t...

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Abstract

A freezing-resistant yeast strain for bread fermentation and a breeding method thereof. The yeast strain provided by the invention is saccharomyces cerevisiae BT-N with an accession number of CGMCC No.7151. When other fermentation performance is not influenced, the strain has a cell freezing survival rate of 92.75% after being frozen at -20 DEG C for 21 days and has a relative fermentation capacity of 53.24%, which are increased by 57.39% and 24.78% respectively when compared with parent strains (who have a freezing survival rate is 35.36% and have a relative fermentation capacity of 28.46%). The strain is realized by knocking out of the neutral trehalase coding gene NTH1 of saccharomyces cerevisiae CICC31616, and overexpression of the trehalose-6-phosphate synthase coding gene TPS1 by selecting a strong promoter PGK1. The bred yeast strain has no special requirements for fermentation equipment or conditions, can be used by equipment and under conditions of common factories, and has wide application prospects.

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 freeze-resistant yeast strain for bread fermentation and a breeding method thereof. Background technique: [0002] Frozen dough technology is a new process of bread production, which is a process of separating the two links of dough making and baking in bread production. Compared with the traditional bread making method, the frozen dough method has the advantages of high production efficiency, stable bread quality, and convenience. However, when the dough is frozen, the temperature is generally 18-20°C, and the yeast in the dough will be seriously damaged during the freezing, freezing and thawing process, especially after the fermented dough goes through the freezing process, the survival rate and gas production of the yeast Ability is significantly reduced. After 20 days of frozen storage of ordinary baker's ye...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81C12R1/865
Inventor 张翠英肖冬光董建吴鸣月孙溪
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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