Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Brewing yeast engineering bacteria containing Lg-ATF1 genes

A technology of Saccharomyces cerevisiae and engineering bacteria, which is applied in the field of bioengineering and can solve the problems of low ester production capacity of Saccharomyces cerevisiae

Active Publication Date: 2012-07-25
河北琢酒集团有限公司
View PDF3 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problem of low ester production ability of Saccharomyces cerevisiae, and provide a Saccharomyces cerevisiae engineering strain containing Lg-ATF1 gene

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Brewing yeast engineering bacteria containing Lg-ATF1 genes
  • Brewing yeast engineering bacteria containing Lg-ATF1 genes
  • Brewing yeast engineering bacteria containing Lg-ATF1 genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: Containing the construction of the Saccharomyces cerevisiae engineered bacteria of Lg-ATF1 gene

[0028] (1) Construction of genetic engineering strains

[0029] 1) Ligate the PGK1 promoter and terminator with the pUC19 plasmid to obtain pUC-PGK1;

[0030] 2) Ligate the homologous fragment IAH derived from Saccharomyces cerevisiae to the pUC-PGK1 obtained in step 1) to obtain pUC-PGK1-IAH;

[0031] 3) Inserting the gene encoding alcohol acetyltransferase Lg-ATF1 in Saccharomyces cerevisiae into the pUC-PGK1-IAH obtained in step 2) between the PGK1 promoter and terminator to obtain pUC-PGK1-IAH-Lg-ATF1;

[0032] 4) connecting the kan gene to the pUC-PGK1-IAH-Lg-ATF1 obtained in step 3) to obtain the plasmid pUC-PGK1-IAH-Lg-ATF1-kan (hereinafter referred to as pUC-PILgK);

[0033] figure 1 The verification electropherogram of the pUC-PILgK plasmid: Lane 1 is the 5000 bp DNA Ladder Marker; Lane 2 is the homologous fragment IAH amplified by PCR from the gen...

Embodiment 2

[0042] Example 2: Research on the fermentation performance of Saccharomyces cerevisiae engineering bacteria and starting strains containing Lg-ATF1 gene

[0043] The engineering bacteria and the recipient bacteria were respectively inserted into 5mL wort culture solution, and cultured overnight at 30°C for 12h; all the bacterial liquids were transferred to 20mL fresh wort culture liquid, and cultured at 30°C for 24h. Take 100g of japonica rice and place it in water at 25-30°C for 72 hours; take it out, wash it, and cook it under normal pressure for 30 minutes. Cool the rice, put it into a 500mL conical flask, add 10g of cooked wheat koji and 105mL of water. Finally, 25 mL of yeast wort culture liquid was inserted and fermented at 28° C. for 5 days. Shake and weigh every 12 hours during the fermentation period, and record the weight loss; after the fermentation is over, stop the cultivation and weigh; measure the residual sugar concentration, alcohol volume fraction and main a...

Embodiment 3

[0047] Example 3: Research on the fermentation performance of engineering haploids containing the Lg-ATF1 gene and the haploids of the starting strain

[0048] Put the engineering haploid (type a / α) and the haploid of the recipient bacteria (type a / α) into 5mL wort culture solution, culture overnight at 30°C for 12h; transfer all the bacteria solution to 20mL fresh wort Incubate at 30°C for 24 hours in the culture medium. Take 100g of japonica rice and place it in water at 25-30°C for 72 hours; take it out, wash it, and cook it under normal pressure for 30 minutes. Cool the rice, put it into a 500mL conical flask, add 10g of cooked wheat koji and 105mL of water. Finally, 25 mL of yeast wort culture liquid was inserted and fermented at 28° C. for 5 days. Shake and weigh every 12 hours during the fermentation period, and record the weight loss; after the fermentation is over, stop the cultivation and weigh; measure the residual sugar concentration, alcohol volume fraction and ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to brewing yeast engineering bacteria containing Lg-ATF1 genes; a method for preparing the yeast engineering bacteria provided by The invention comprises the following steps: selecting strong promoters PGK1 so as to over express specific Lg-ATF1 genes coding alcohol acetyl transferase of beer yeast in brewing yeast, thereby obtaining the yeast engineering bacteria SaccharomycescerevisiaeEY-15 containing Lg-ATF1 genes, wherein a preservation number is CGMCC (China General Microbiological Culture Collection Center) No.5635. According to The invention, on a condition that other fermenting performance is not influenced, transformant bacterial strains are compared with parent strains (SaccharomycescerevisiaeCGMCC No.1525); after simulation of semi-solid fermentation, the content of ethyl acetate is increased by 1.6 times; the content of isoamyl acetate is increased to 26.6mg / L; the sifted engineering bacteria has no specific demand for fermentation equipment and conditions; the equipment and conditions of normal white sprits factories can be used; therefore, the engineering bacteria has wide application propose and brings marked economic benefit to industrial production of the white sprits factories.

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 engineering bacteria containing Lg-ATF1 gene. Background technique [0002] How to increase the content of ester aroma substances in wine has always been an important subject of research by my country's ordinary liquor, yellow rice wine enterprises and related scientific research units. At present, there are three main methods for increasing the ester content of ordinary liquor: the one is the solid-liquid combination method, which uses the liquid method to produce the wine base, and uses the distiller's grains, tails or finished wine of the solid method to improve the quality; Modulate natural spices or use pure chemicals to flavor according to the aroma components of a certain famous wine; the third is the whole liquid method, adding aroma-producing microorganisms, caproic acid ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/04C12R1/865
Inventor 肖冬光张翠英郭学武张建炜戴隆海
Owner 河北琢酒集团有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com