Genetically engineered bacterium with low yield of 2-phenethyl alcohol and application of genetically engineered bacterium

A genetically engineered bacterium and gene technology, applied in the field of Saccharomyces cerevisiae genetically engineered bacteria with low production of 2-phenylethanol, can solve the problems of high 2-phenylethanol content and low production of 2-phenylethanol, etc., to reduce 2-phenylethanol, improve Good flavor, fermentation performance and growth performance

Active Publication Date: 2020-12-01
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • 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 that Saccharomyces cerevisiae strains produce 2-phenylethanol content higher in the production of liquo

Method used

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  • Genetically engineered bacterium with low yield of 2-phenethyl alcohol and application of genetically engineered bacterium
  • Genetically engineered bacterium with low yield of 2-phenethyl alcohol and application of genetically engineered bacterium
  • Genetically engineered bacterium with low yield of 2-phenethyl alcohol and application of genetically engineered bacterium

Examples

Experimental program
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Effect test

Example Embodiment

[0031] Example 1: Gene deletion ARO80 Construction of S. cerevisiae strains

[0032] Saccharomyces cerevisiae α5 (Li, W., Wang, JH, Zhang, CY, Ma, HX, & Xiao, DG (2017) .Regulation of Saccharomyces cerevisiae genetic engineering on the productionof acetate esters and higher alcohols during Chinese Baijiufermentation.Journal of industrial microbiology & biotechnology, 44 (6), 949-960.https: / / doi.org / 10.1007 / s10295-017-1907-2) strain as a host to construct a recombinant genetic engineering strain ARO80 deleted gene by homologous recombination.

[0033] Specific construction steps detailed below:

[0034] (1) genome starting strain Saccharomyces cerevisiae α5 as a template to ARO80A-F and ARO80A-R as primers by PCR amplification of upstream DNA molecule fragments ARO80A (1779bp) obtained ARO80 gene; Saccharomyces cerevisiae α5 genome template, in ARO80B-F and ARO80B-R was amplified using PCR primers downstream DNA molecule fragments ARO80B (1758bp) obtained ARO80 gene.

[0035] (2) ...

Example Embodiment

[0046] Example 2: recombinant strain α5-ΔARO80-k-p fermentation experiment liquid fermentation liquor

[0047] (1) fermentation process route:

[0048] → pulverization → sorghum particles liquefaction, saccharification addition of an acidic protease → → → filtration → cooling sorghum sugar juice dispensing → Adjustment → sterilization → inoculation, fermentation → distillation;

[0049] (2) the main conditions are as follows:

[0050] Pulverization conditions: no grinding of whole sorghum is appropriate, the degree of pulverization is not easy too small to avoid excessive pressure filtration;

[0051] Liquefaction, saccharification conditions: Sorghum pulverized to water ratio 1: 4 ratio of 30 deg.] C warm water was added, and after stirring sufficiently homogeneous, placed in a constant temperature water bath, kept 60min 90 ℃, liquefaction. Adjusting the water bath temperature to 60 ℃, kept 30min, saccharification. 5min sufficiently stirred once every liquefaction and saccharific...

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Abstract

The invention provides a genetically engineered bacterium with low yield of 2-phenethyl alcohol. The genetically engineered bacterium is obtained by completely deleting ARO80 genes through saccharomyces cerevisiae. A transcription factor Aro80p encoded by the ARO80 gene activates the transcription of an aromatic amino acid catabolic gene in the presence of aromatic amino acids, and promotes the synthesis of amino acid aminotransferase I and aromatic amino acid aminotransferase II. The deletion of the ARO80 gene hinders the activation of the expression of the ARO9 and ARO10 genes, thereby regulating the synthesis of the 2-phenethyl alcohol in the Ehrlich pathway and obtaining the saccharomyces cerevisiae strain with low yield of the 2-phenethyl alcohol. The genetically engineered bacteriumdisclosed by the invention is applied to production of white spirit by a liquid fermentation method.

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 Saccharomyces cerevisiae genetically engineered bacterium with low 2-phenylethanol production and its application. Background technique [0002] The flavor substances in liquor mainly include higher alcohols, esters, and aldehydes. Among them, higher alcohol is one of the important chemical substances that form the flavor and mouthfeel of liquor. Appropriate higher alcohol content has the effect of making the taste and aroma of the liquor plump, soft and harmonious, but excessive higher alcohol is the main source of the off-flavor of the liquor, and it is also an important reason for the baijiu to swell after drinking. As one of the important components in higher alcohols, 2-phenylethanol has a great influence on the style of liquor. An appropriate amount of 2-phenylethanol can bring rose aroma to the body of the ...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12N15/90C12G3/021C12R1/865
CPCC07K14/395C12N15/905C12G3/021
Inventor 肖冬光王亚平张翠英陈叶福杜丽平郭学武
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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