Acetic acid resistant ethanol producing wine making yeast strains and strain screening method

A technology of Saccharomyces cerevisiae strain and acetic acid tolerance, which is applied in the field of genetic breeding of industrial microorganisms, can solve the problems of loss of fermentable sugar, increase of production cost, reduction of inhibitor concentration, etc. simple effect

Inactive Publication Date: 2012-12-12
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Water washing detoxification pretreatment can reduce the concentration of inhibitors, but at the same time, a large amount of fermentable sugar will be lost, which will increase the production cost

Method used

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  • Acetic acid resistant ethanol producing wine making yeast strains and strain screening method
  • Acetic acid resistant ethanol producing wine making yeast strains and strain screening method
  • Acetic acid resistant ethanol producing wine making yeast strains and strain screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Screening of acetic acid tolerant Saccharomyces cerevisiae E5.9

[0041] 1) Starting strain

[0042] By analyzing the stress tolerance of 96 Saccharomyces cerevisiae strains from different sources, the strain XL14-1 with strong comprehensive stress resistance was selected as the starting strain. The strains were identified by phenotype and ribosomal gene 26S rDNA D1 / D2 region and ITS region sequence analysis.

[0043] 2) Straw pretreatment fermentation broth

[0044] The solid matter of straw steam explosion pretreatment was provided by COFCO Biochemical Energy (Zhaodong) Co., Ltd., and the hydrolyzate was obtained after repeated extraction. Add glucose 15%, yeast extract 0.6%, peptone 1%, (NH 4 ) 2 SO 4 0.2%, KH 2 PO 4 0.1%, MgSO 4 .7H 2 O 0.15%, CaCl 2 0.055% is made into a pretreatment fermentation liquid, and the final concentration of glucose is 20.5%.

[0045] 3) Mutagenesis

[0046] The three mutagenesis methods adopted in the present inv...

Embodiment 2

[0067] Embodiment 2: Stress resistance screening method - liquid full screening method

[0068] 1. Liquid full screening process

[0069] The method comprises four steps: (1) strain mutagenesis, (2) liquid full screening, (3) solid plate screening, (4) subculture and domestication, wherein liquid full screening is the core of the method, see Figure 5 . First, use ultraviolet, EMS, microwave or other mutagenesis methods to mutate Saccharomyces cerevisiae cells to obtain a library of mutant strains; secondly, transfer all the cells after mutagenesis treatment to a 24-well plate on average, 120 rpm, 30°C pre- Cultivate for 6 hours, then add selective pressure to the pre-cultured bacteria solution, and continue to cultivate until positive mutations of resistance appear; again, dilute the positive mutant bacteria solution and spread the same concentration of solid selection medium, and when colonies grow out, pick large The colony was cultured in the same pressure liquid medium,...

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Abstract

The invention discloses acetic acid resistant ethanol producing wine making yeast strains, which is named Saccharomyces cerevisiae) E5.9 in taxonomy and has a collection number of CGMCC N0.3618. A screening method, which is a liquid full-screening method, comprises the following steps: 1) strain mutation induction, namely performing the physical, chemical or biological mutation induction of starting stains of the wine making yeasts; 2) liquid full-screening, namely uniformly transferring the strains subjected to mutation induction into a 24-hole plate, culturing under a selective pressure-free condition, and continuing to culture under a selective pressure till positive mutants are generated in a plurality holes; 3) solid flat plate screening, namely diluting a positive mutation culture material, coating the positive mutation culture medium on a solid selective culture medium, and selecting an advantageous resistant clone; and 4) filial generation domestication, namely performing the filial generation domestication of the advantageous resistant clone in a selective liquid culture medium, and selecting stress resistance strains with genetic stability.

Description

technical field [0001] The invention belongs to the field of genetic breeding of industrial microorganisms, and in particular relates to an acetic acid-tolerant ethanol-producing Saccharomyces cerevisiae strain E5.9 and a screening method. Background technique [0002] Lignocellulose is the most abundant renewable biomass resource, but it has not been well utilized so far. Fuel ethanol is a renewable and clean energy that is expected to become one of the substitutes for fossil energy and is beneficial to environmental protection. The production of fuel ethanol from lignocellulose is of great economic and social significance. [0003] The production of fuel ethanol from lignocellulose requires pretreatment of the raw material. At present, various pretreatment methods such as steam explosion and dilute acid have been developed, but no matter which method is used, there are inevitably fermentation inhibitors in the pretreatment, which seriously affects the growth of fermentati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/18C12N15/01C12P7/10C12R1/865
CPCY02E50/16Y02E50/10
Inventor 王士安袁博李福利
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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