Industrial saccharomyces cerevisiae strain for synchronously producing xylitol and ethanol and construction method of industrial saccharomyces cerevisiae strain

The technology of a Saccharomyces cerevisiae strain and its construction method is applied in the field of industrial Saccharomyces cerevisiae strains and its construction, which can solve the problems of unsatisfactory fermentation requirements, slow xylose metabolism rate, and low xylitol production, and achieve xylose consumption rate acceleration, The effect of increasing xylose consumption rate

Pending Publication Date: 2022-03-04
CHINA PETROLEUM & CHEM CORP +2
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  • Abstract
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Problems solved by technology

[0005] The object of the present invention is to solve the problem that after the existing CtXYL1 (encoding xylose reductase) gene derived from Candida tropicalis is expressed heterologously in Saccharomyces cerevisiae, in order to simulate its fermentation process in lignocellulose hydrolyzate, select a high initial concentration When glucose is supplied as a carbon source, the recombinant bacteria constructed by the xylose metabolism rate is slow, the xylitol yield is low, and cannot meet the needs of fermentation, and then a high-expression FPS1 gene is provided to solve the above problems, and the SOR1 / SOR2 The industrial Saccharomyces cerevisiae strain SEB20 with high xylitol and ethanol production under co-fermentation glucose and xylose conditions by replacing the gene coding region with CtXYL1 gene and its construction method

Method used

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  • Industrial saccharomyces cerevisiae strain for synchronously producing xylitol and ethanol and construction method of industrial saccharomyces cerevisiae strain
  • Industrial saccharomyces cerevisiae strain for synchronously producing xylitol and ethanol and construction method of industrial saccharomyces cerevisiae strain
  • Industrial saccharomyces cerevisiae strain for synchronously producing xylitol and ethanol and construction method of industrial saccharomyces cerevisiae strain

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Embodiment 1

[0050] The present embodiment provides a kind of industrial saccharomyces cerevisiae strain of synchronous production xylitol and ethanol, and its construction method comprises the following steps:

[0051] (1) Introducing the Cas9-NAT plasmid into Saccharomyces cerevisiae SEB10: the existing plasmid Cas9-NAT (1 μg) (the nucleic acid sequence is SEQ ID NO.1) was transformed into the Saccharomyces cerevisiae SEB10 strain by the lithium acetate transformation method. Positive transformants with good colony morphology were screened in the 2% YPD plate of mycetin;

[0052] (2) Crude extraction of genomic DNA from Saccharomyces cerevisiae transformants: pick possible positive transformants into 95 μL 1% SDS (sodium dodecylsulfonate) with a toothpick, and add 4 μL 4M lithium acetate solution, and then follow the published steps In a document reported by Robert Mans et al. in 2015, the title of the document is: CRISPR / Cas9: a molecular Swissarmy knife for simultaneous introduction of...

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Abstract

The invention provides an industrial saccharomyces cerevisiae strain for synchronously producing xylitol and ethanol and a construction method of the industrial saccharomyces cerevisiae strain. The strain highly expresses an FPS1 gene and an XYL1 gene. Specifically, the strain is SEB20, the classification name of the strain is Saccharomyces cerevisiae, and the preservation number of the strain is CGMCC No.22590. The strain is obtained by taking the Saccharomyces cerevisiae SEB10 as an original strain, replacing a promoter PFPS1 of a gene FPS1 with a strong promoter PTDH3, and increasing the copy number of a CtXYL1 gene while knocking out two site gene coding regions of SOR1 and SOR2. Under the condition that the initial glucose concentration of the saccharomyces cerevisiae strain SEB20 is relatively high, the xylose consumption rate and the yield of xylitol and ethanol are improved. Under a test tube fermentation condition, compared with SEB10, the xylose consumption rate is increased by 73.5%, and the xylitol yield is increased by 27.0%. Under the conditions of straw pre-saccharification and simultaneous saccharification fermentation, compared with an original strain SEB10, the strain SEB20 has the advantages that the xylose consumption rate is increased by 24.1%, the xylitol yield is increased by 42.9%, the xylitol yield is close to a theoretical value 1.0, and the ethanol yield is increased by 5.0%.

Description

technical field [0001] The invention relates to the technical field of microbial genetic engineering, in particular to an industrial Saccharomyces cerevisiae strain synchronously producing xylitol and ethanol and a construction method thereof. Background technique [0002] The massive consumption of fossil fuels has caused serious problems such as energy depletion, rising prices and global climate change. Lignocellulosic biomass such as agricultural waste (bagasse, corncobs, corn stalks, wheat and rice straw, etc.), waste wood, and industrial waste (including pulp, paper processing waste, etc.) can be used as potential raw materials for biofuels. It is widely sourced, low-cost, and undergoes environmentally sound biological treatment to produce fuel ethanol, which benefits rural communities and society as a whole with macroeconomic benefits. In China, more than 900 million tons of straw are produced every year, and the comprehensive utilization rate of straw exceeds 85%. Af...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81C12P7/18C12P7/06C12R1/865
CPCC07K14/395C12N9/0006C12N15/81C12Y101/01307C12P7/18C12P7/06Y02E50/10
Inventor 汤岳琴陈栋杨白雪谢采芸
Owner CHINA PETROLEUM & CHEM CORP
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