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Genetically engineered bacterium with low n-propanol yield and application thereof

A technology of genetically engineered bacteria and genes, applied in the direction of genetic engineering, application, plant genetic improvement, etc., can solve the problems of low yield and high n-propanol content, improve flavor, reduce n-propanol, and have good fermentation performance and growth performance. Effect

Active Publication Date: 2020-12-01
TIANJIN UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem of higher content of n-propanol produced by Saccharomyces cerevisiae strains in the production of liquor by the liquid method, and to construct Saccharomyces cerevisiae strains with low production of n-propanol by regulating the threonine synthesis pathway of Saccharomyces cerevisiae

Method used

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  • Genetically engineered bacterium with low n-propanol yield and application thereof
  • Genetically engineered bacterium with low n-propanol yield and application thereof
  • Genetically engineered bacterium with low n-propanol yield and application thereof

Examples

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Effect test

Embodiment 1

[0030] Example 1: Construction of Saccharomyces cerevisiae Strains Deleting the THR6 Gene

[0031] Saccharomyces cerevisiae α5 (Li W, Wang JH, Zhang CY, Ma HX, Xiao DG (2017b) Regulation of Saccharomyces cerevisiae genetic engineering on the production of acetateesters and higher alcohols during Chinese Baijiu fermentation. J Ind Microbiol Biotechnol 44:949-960.) As the host bacteria, recombinant genetically engineered strains are constructed by means of homologous recombination.

[0032] The specific construction steps are detailed as follows:

[0033] (1) Using the genome of the starting strain Saccharomyces cerevisiae α5 as a template, using THR6A-F and THR6A-R as primers, amplify the upstream DNA molecular fragment THR6A (972bp) of the THR6 gene by PCR; use the Saccharomyces cerevisiae α5 genome as As a template, THR6B (724bp), a downstream DNA molecule fragment of the THR6 gene, was amplified by PCR using THR6B-F and THR6B-R as primers.

[0034] (2) Using the plasmid pU...

Embodiment 2

[0045] Example 2: Liquor fermentation experiment of recombinant strain α5-ΔTHR6-k-p liquid method

[0046] (1) Fermentation process route:

[0047] Sorghum granules→crushing→liquefaction, saccharification→adding acid protease→cooling→filtering→adjustment of sugar content of sorghum juice→subpackaging→sterilization→inoculation, fermentation→distillation;

[0048] (2) The main process conditions are as follows:

[0049] 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;

[0050] 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 n-propanol yield. The genetically engineered bacterium is obtained by completely deleting a homoserine dehydrogenase gene THR6 from saccharomyces cerevisiae, wherein the homoserine dehydrogenase encoded by the THR6 gene can catalyze dimer enzyme in a third step of a threonine biosynthetic pathway, and the dimer enzyme plays a key role in the synthetic pathway of saccharomyces cerevisiae threonine. Threonine can be converted into 2-ketobutyric acid. 2-ketobutyric acid is a necessary precursor substance for synthesizing n-propanol by saccharomyces cerevisiae, and directly influences the yield of n-propanol. The genetically engineered bacterium disclosed 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 bacteria with low n-propanol 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, n-propanol has a greater impact on the style of liquor. An appropriate amount of n-propanol can bring wine aroma and sweetness to the body of the liq...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/87C12N15/81C12N15/53C12G3/02C12G3/021C12R1/865
CPCC12N9/0006C12N15/87C12N15/81C12G3/02C12G3/021C12Y101/01003C12N2800/30Y02E50/10
Inventor 肖冬光王亚平张翠英陈叶福杜丽平郭学武
Owner TIANJIN UNIV OF SCI & TECH
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