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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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  • Abstract
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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 liquor by liquid method, and produce 2-phenylethanol by regulating Saccharomyces cerevisiae Construction of Saccharomyces cerevisiae strains with low 2-phenylethanol production

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

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Embodiment 1

[0031] Example 1: Construction of Saccharomyces cerevisiae Strains Deleting the ARO80 Gene

[0032] Taking Saccharomyces cerevisiae α5 (Li, W., Wang, J.H., Zhang, C.Y., Ma, H.X., & Xiao, D.G. (2017). Regulation of Saccharomyces cerevisiae genetic engineering on the production of 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) was used as the host bacteria, and a recombinant genetically engineered strain lacking the ARO80 gene was constructed by homologous recombination.

[0033] The specific construction steps are detailed as follows:

[0034] (1) Using the genome of the starting strain Saccharomyces cerevisiae α5 as a template, using ARO80A-F and ARO80A-R as primers, the upstream DNA molecular fragment ARO80A (1779bp) of the ARO80 gene was obtained by PCR amplification; using the Saccharomyces cerevisiae α5 genome as a template, Using ARO80B-F an...

Embodiment 2

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

[0047] (1) Fermentation process route:

[0048] Sorghum granules→crushing→liquefaction, saccharification→adding acid protease→cooling→filtering→adjustment of sugar content of sorghum juice→subpackaging→sterilization→inoculation, fermentation→distillation;

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

[0050] Grinding conditions: the degree of crushing is suitable for sorghum without whole grains, and the degree of crushing should not be too fine, so as not to cause excessive filtration pressure;

[0051] Liquefaction and saccharification conditions: add crushed sorghum to warm water at 30°C at a material-to-water ratio of 1:4, stir well, place in a constant temperature water bath, keep at 90°C for 60 minutes, and liquefy. Adjust the temperature of the water bath to 60°C and keep it for 30 minutes for saccharification. Fully stir once every 5 minutes during liquefa...

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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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