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Industrial saccharomyces cerevisiae strain capable of producing xylitol and construction method of industrial saccharomyces cerevisiae strain

A technology of Saccharomyces cerevisiae strain and construction method, which is applied to the field of industrial Saccharomyces cerevisiae producing xylitol and construction, and can solve the problems of low expression level, high xylitol yield, loss of exogenous genes and the like

Active Publication Date: 2016-03-02
SICHUAN UNIV
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Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide an industrial Saccharomyces cerevisiae strain producing xylitol, which not only overcomes the problems of exogenous gene loss and low expression level during the expression of existing Saccharomyces cerevisiae strains; high

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  • Industrial saccharomyces cerevisiae strain capable of producing xylitol and construction method of industrial saccharomyces cerevisiae strain
  • Industrial saccharomyces cerevisiae strain capable of producing xylitol and construction method of industrial saccharomyces cerevisiae strain
  • Industrial saccharomyces cerevisiae strain capable of producing xylitol and construction method of industrial saccharomyces cerevisiae strain

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Embodiment

[0047] A method for constructing a xylitol-producing industrial Saccharomyces cerevisiae strain, comprising the following steps: (1), designing and amplifying primers containing a target gene: according to the published δ sequence in the NCBI database, designing and synthesizing two pairs of primers: one of A pair of primers is δ-pB-F (SEQ ID NO.2), δ-pB-R (SEQ ID NO.3); another pair of primers is δ-XR-XDH-F (SEQ ID NO.4), δ-pB-R (SEQ ID NO. .3);

[0048] (2) Preparation of pK-XR-Ct plasmid;

[0049] (3) Amplification of the target gene: using the pK-XR-Ct plasmid as a template and δ-pB-F and δ-pB-R as primers, amplify xyl1 containing xylose reductase gene and G418 resistance screening gene Linearized DNA fragment A of KanMX, and purify and recover fragment A obtained by PCR amplification; the promoter of gene xyl1 is pGAPDH, and the terminator is tGAPDH; the promoter of KanMX gene is pTEF1, and the terminator is tTEF1; PCR amplification reaction system (50 μl) is composed a...

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Abstract

The invention relates to an industrial saccharomyces cerevisiae strain capable of producing xylitol and a construction method of the industrial saccharomyces cerevisiae strain. The preservation number of the saccharomyces cerevisiae strain is CGMCC 11326. The construction method comprises the following steps: taking pK-XR-Ct plasmids as a template, taking delta-pB-F and delta-pB-R as primers, and carrying out PCR amplification to obtain DNA fragments A, taking the DNA fragments A as a template, and carrying out amplification to obtain DNA fragments B; screening the G418 optimal tolerance concentration of a saccharomyces cerevisiae fusion strain; converting the saccharomyces cerevisiae fusion strain; and screening unicellular converters to obtain the saccharomyces cerevisiae strain. The saccharomyces cerevisiae SEB6 strain prepared by the construction method provided by the invention has excellent performances of producing xylitol through xylose fermentation, and the yield of xylitol is high; and meanwhile, the problems that exogenous genes get lost and the expression level is low existing in the expression of the existing saccharomyces cerevisiae strains are avoided.

Description

technical field [0001] The invention relates to the technical field of microbial genetic engineering, in particular to a xylitol-producing industrial Saccharomyces cerevisiae strain and a construction method. Background technique [0002] Agricultural wastes such as straws, husks, and cores are rich in crude fibers (pentanose), but these agricultural wastes are mostly disposed of by waste or incineration, resulting in a great waste of resources and environmental pollution. And xylitol can be obtained by hydrolysis of crude fiber (pentose). Using agricultural wastes as raw materials to prepare xylitol is an effective way to turn agricultural and sideline products into treasures, protect the environment, and save resources. As a functional sweetener, xylitol is an adjuvant therapeutic agent for diabetics. It has functions such as weight loss, intestinal regulation, and liver function improvement. It is widely used in the food industry. [0003] The methods for producing xyli...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81C12R1/865
Inventor 汤岳琴杨白雪苟敏孙照勇木田建次夏子渊
Owner SICHUAN UNIV
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