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High-yield ethyl lactate saccharomyces cerevisiae strain

A technology of ethyl lactate and Saccharomyces cerevisiae, which is applied in the field of bioengineering, can solve the problem of low ability to synthesize ethyl lactate, achieve the effect of increasing the production amount and increasing ethyl lactate

Inactive Publication Date: 2019-04-19
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that Saccharomyces cerevisiae has a low ability to synthesize ethyl lactate in wine production, and provide a high-yielding Saccharomyces cerevisiae strain

Method used

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  • High-yield ethyl lactate saccharomyces cerevisiae strain
  • High-yield ethyl lactate saccharomyces cerevisiae strain
  • High-yield ethyl lactate saccharomyces cerevisiae strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: Construction of high-yield ethyl lactate Saccharomyces cerevisiae strain

[0031] The main construction process of the strain is as follows:

[0032] Yip-rPCk was constructed based on the plasmid Yip-lac211, see the plasmid construction procedure figure 1. First, the α5 genome of the haploid strain of Saccharomyces cerevisiae CICC32315 was used as a template, and the rDNA (SEQNO: 2) fragment, 2269bp, was amplified by PCR using primers rDNA-U and rDNA-D. The laboratory-preserved plasmid Yep-PGK was used as a template, and PGK-U and PGK-D were used as primers to amplify the PGK1 fragment with a length of 1771 bp. The rDNA and PGK1 were connected together by fusion PCR to obtain PGK1-rDNA, and were ligated to the plasmid Yip-lac211 by SacI and NcoI to obtain Yip-rP. Using the Yep-PCnK plasmid as a template, primers CALB-U and CALB-D, the target fragment CALBn (SEQ NO: 1) was obtained by PCR, which was ligated to Yip-rP using BglII single enzyme digestion t...

Embodiment 2

[0039] Example 2: Fermentation experiment of multi-copy strain sorghum raw material semi-solid liquor

[0040] 1) Fermentation process roadmap:

[0041] Sorghum → soaking → liquefaction → saccharification → cooling → mixing koji, inoculation with lactic acid bacteria → cultivation for 24 hours → fermentation → distillation

[0042] 2) Process conditions: soaking conditions: 60-70°C, fully absorb water; cooking conditions: 121°C for about 30 minutes; liquefaction conditions: 85-90°C, high temperature resistant α-amylase, liquefaction for 60 minutes; saccharification conditions: 55-60°C , add glucoamylase, saccharification for 30min; fermentation conditions: 30°C, 6 days; wine steaming conditions: 100mL fermentation broth, add 100mL water, steam 100mL wine sample.

[0043] 3) Ingredients: sorghum flour: 78g; add water 200mL; high temperature resistant α-amylase: 25μL; glucoamylase: 45μL; Daqu 12g;

[0044] Carry out semi-solid Daqu liquor fermentation experiments on Saccharomy...

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Abstract

The present invention provides a high-yield ethyl lactate saccharomyces cerevisiae strain, which includes the construction of a multi-copy integration carrier and fermentation validation of the selected strain. The CALB gene is taken as a target gene, the selected yeast strain performs multi-copy expression of Pseudozyma antarctica lipase B, compared with the parent strain, the basic fermentationperformance of the constructed Saccharomyces cerevisiae strain is not affected, after 6 days of fermentation of sorghum raw material semi-solid white wine, the content of ethyl lactate in the selectedstrain is 223.159 mg / L, the amount of ethyl lactate in the parent strain is 114.04 mg / L, and the multi-copy strain is increased by 92.3% compared to the parent strain. The selected Saccharomyces cerevisiae strain significantly increases the amount of ethyl lactate, makes the liquor more abundant and more harmonious in taste, and has a wide range of applications in the field of liquor.

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 Saccharomyces cerevisiae with high yield of ethyl lactate. Background technique [0002] The content of trace flavor substances in liquor accounts for about 1%-2% of liquor, and is the main factor affecting the quality of liquor, while esters account for 60%-90% of trace flavor substances, so increasing the ester content of liquor is important It is very important to enrich the taste of liquor and improve the coordination of liquor. The content of ethyl lactate in the ester compounds of Fen-flavor liquor is second only to ethyl acetate, and the content of ethyl lactate has a great influence on the flavor of liquor. Due to the gradual realization of comprehensive mechanization of modern large-scale liquor enterprises and the gradual improvement of sanitation and cleanliness, the amount of ethyl lactate pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81C12G3/021C12R1/865
CPCC12N9/20C12G3/02C12N15/81C12Y301/01003
Inventor 张翠英肖冬光刘鑫葛峻伶郭学武陈叶福董建杜丽平马丽娟
Owner TIANJIN UNIV OF SCI & TECH
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