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Genetic recombination saccharomyces cerevisiae with detoxification function and construction method and application thereof

A technology of Saccharomyces cerevisiae and gene recombination, applied in the field of genetic engineering, can solve the problems of low utilization rate of xylose, infeasible detoxification process in production, inability to remove inhibitors and the like

Active Publication Date: 2019-11-15
ZHONGRONG TECH CORP LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, physical and chemical detoxification methods have their insurmountable shortcomings: a single method cannot remove inhibitors with different physical and chemical properties; the combined use of multiple physical and chemical methods will greatly increase the cost of detoxification, and complex detoxification processes become unfeasible in production ; Some physical and chemical detoxification methods will also produce new pollutants
However, when this strain ferments lignocellulose hydrolyzate, the utilization rate of xylose is very low. The main reason is that various inhibitors in the hydrolyzate inhibit the utilization of xylose by this strain.

Method used

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  • Genetic recombination saccharomyces cerevisiae with detoxification function and construction method and application thereof
  • Genetic recombination saccharomyces cerevisiae with detoxification function and construction method and application thereof
  • Genetic recombination saccharomyces cerevisiae with detoxification function and construction method and application thereof

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Embodiment Construction

[0041] In order to better explain the present invention and facilitate understanding, the present invention will be described in detail below through specific embodiments in conjunction with the accompanying drawings.

[0042] In order to obtain a kind of gene recombinant Saccharomyces cerevisiae with detoxification function, the following method can be carried out:

[0043] (1) Extraction of Pichia stipitis genomic DNA

[0044] The Pichia stipitis genomic DNA was extracted using the Yeast Genomic DNA Extraction Kit of Solebo Company, and verified by 1% agarose gel electrophoresis after the extraction was completed. The specific operation steps are as follows:

[0045] (1) Take 1-5ml of yeast cells, centrifuge at 12000rpm for 1min, and remove the supernatant as much as possible.

[0046] (2) Add 470ul sorbitol Buffer to the yeast cells. Fully suspend the bacteria, add 25ul yeast wall-breaking enzyme and 5ul sulfhydryl reducing agent, and mix well. Treat at 30°C for 1-2 hour...

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Abstract

The invention relates to genetic recombination saccharomyces cerevisiae with a detoxification function. The genetic recombination saccharomyces cerevisiae contains an exogenous aldehyde reductase gene, and the exogenous aldehyde reductase gene is an aldehyde reductase gene of which the sequence is shown in SEQ ID NO:1. The exogenous aldehyde reductase gene is obtained in a way that genome DNA of pichia stipitis is taken as a template for PCR amplification, and meanwhile, two enzyme digestion sites including Xho I and Not I are introduced into the terminal C and the terminal N respectively. A GRE2 gene segment is connected to a pYES2 / NTA plasmid vector to obtain a recombinant plasmid pYES2 / NTA-GRE2, and then the recombinant plasmid pYES2 / NTA-GRE2 is transformed into the saccharomyces cerevisiae to obtain the genetic recombination saccharomyces cerevisiae with the detoxification function. The saccharomyces cerevisiae can efficiently ferment pretreatment materials of lignocellulose to produce fuel ethanol; meanwhile, inhibitory or toxic aldehyde substances generated in the fermentation process can be removed, and detoxification of fermentation inhibiting substances in the pretreatmentmaterials of the lignocellulose is achieved, so that it is ensured that the process of producing the ethanol through fermentation of the cellulose is smooth and efficient.

Description

technical field [0001] The invention relates to genetic engineering technology, in particular to a gene recombinant Saccharomyces cerevisiae with detoxification function and its construction method and application. Background technique [0002] With the increasing shortage of petroleum resources and declining reserves, using agricultural straw as raw material to ferment and produce fuel ethanol and various chemical products to meet the needs of human social development is considered to be a clean energy that can replace petroleum. Experts believe that the production of ethanol from lignocellulosic biomass represented by crop straws has good development prospects, and at the same time helps to solve the problem of agricultural production waste disposal, which not only alleviates the resource crisis and food crisis, but also has a great impact on environmental pollution. It is of great significance and provides a guarantee for more sustainable development. [0003] Lignocellu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/53C12N15/81C12N15/66C12P7/10C12R1/865
CPCC12N9/0006C12N15/66C12N15/81C12P7/10C12Y101/01002Y02E50/10
Inventor 杨何宝李秋园代淑梅
Owner ZHONGRONG TECH CORP LTD
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