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A highly tolerant yeast strain and its construction method

A yeast strain, high-tolerance technology, applied in the field of bioengineering, can solve problems such as blanks, achieve good resistance to high sugar, high cell viability, and improve the level of production technology.

Active Publication Date: 2017-12-19
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At the same time, the current research on H / ACA small nucleolar RNA is basically focused on theoretical research, and there are only a few reports on the application of H / ACA small nucleolar RNA, especially in the selection of resistant strains.

Method used

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  • A highly tolerant yeast strain and its construction method
  • A highly tolerant yeast strain and its construction method
  • A highly tolerant yeast strain and its construction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Construction of High Temperature Resistance, High Sugar Resistance, Freezing Resistance Baker's Yeast Strain

[0038] (1) Construction of recombinant plasmid Yep-KPS

[0039] The construction process of the recombinant plasmid Yep-KPS is as follows: figure 1 shown;

[0040]Using the total DNA of yeast strain CICC31616 as a template, PCR amplified the complete sequence of the SNR84 gene;

[0041] Upstream primer S1: 5'-GGA AGATCT ATTGCACAACTTAAGTTTGTCGAGG-3' (SEQ ID NO: 2);

[0042] Downstream primer S2: 5'-GGA AGATCT TAATGTGTCTCTTTGAGTCATGTTCCTT-3' (SEQ ID NO: 3); the underlined part is the restriction site;

[0043] PCR reaction conditions: 95°C for 5min; 94°C for 40s, 54°C for 1min, 72°C for 40s, 30 cycles; 72°C for 10min, 0.8% agarose gel electrophoresis to identify the amplified product;

[0044] The PCR product was connected to the pUC-PGK1 vector containing a strong promoter to obtain pUC-PGKS; using pUC-PGKS as a template, PCR amplification obt...

Embodiment 2

[0053] Example 2: Fermentation experiment of high temperature resistant, high sugar resistant, and freezing resistant baker's yeast strains

[0054] (1) High temperature tolerance experiment of recombinant strains and starting strains

[0055] A ring of yeast cells was picked and cultured in YEPD liquid medium to exponential phase (OD 660 =1–1.5); cells were transferred into 5mL fresh YEPD liquid medium, and the cell OD was adjusted with YEPD liquid medium 600 After = 1.0, take 200 μL of cell culture solution into a 1.5 mL centrifuge tube, heat shock in a water bath at 52°C for 2 min, and immediately ice-bath. After the cells are cooled, take 50 μL of the cell culture and dilute to a certain concentration, and at the same time, dilute the unheated yeast cells to the same multiple, spread them on the YEPD plate and culture them for 2 days, count the number of single colonies, and record them as u 1 , u 2 . Cell survival rate at high temperature (%) = (u 1 / u 2 )×100%.

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Abstract

The invention discloses a high-resistance yeast strain resistant to high temperature, high sugar and freezing and a construction method thereof. The high-resistance yeast strain selects a strong promoter PGK1 to overexpress the H / ACA small nucleus in the parent yeast strain Kernel RNA (SNR84) full sequence to obtain the high tolerance yeast strain. Compared with the parental strain, the cell activity of the strain is significantly improved in high temperature, high sugar and freezing environments. The baker's yeast has strong resistance to high temperature in heat shock treatment, strong resistance to high sugar in high-sugar dough, and strong resistance to freezing in frozen dough, which solves the technical obstacles and quality defects in the bread making process, and has a wide range of 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 yeast strain with high temperature resistance, high sugar resistance and freezing resistance and a construction method thereof. Background technique: [0002] Baker's yeast is an essential microbial leavening agent and loosening agent in the pasta making process, and yeast is also a biological nutrient that can increase the nutritional content of dough. With the development of the baking industry, various dough technologies have emerged as the times require, and good yeast varieties are an important guarantee for bread quality. However, during the baking process of bread, baker's yeast is often exposed to high temperature, high sugar, freezing and other environments, and the poor growth environment will lead to a decline in the activity of baker's yeast, thereby resulting in a decline in the quality of bread. Bak...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/19C12N15/63A21D8/04C12R1/865
Inventor 张翠英肖冬光林雪董健柏晓雯宋海岩
Owner TIANJIN UNIV OF SCI & TECH
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