Saccharomyces cerevisiae engineering bacterium and its application in production of ethanol

A technology of Saccharomyces cerevisiae and ethanol, which is applied in the biological field and can solve problems such as limiting the industrialization of cellulosic ethanol

Active Publication Date: 2012-10-17
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These factors have greatly limited the process of cellulosic ethanol industrialization

Method used

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  • Saccharomyces cerevisiae engineering bacterium and its application in production of ethanol
  • Saccharomyces cerevisiae engineering bacterium and its application in production of ethanol
  • Saccharomyces cerevisiae engineering bacterium and its application in production of ethanol

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Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1, the acquisition of recombinant bacterial strain W32N55

[0031] 1. Construction of Recombinant Engineering Bacteria

[0032] 1. Plasmid construction

[0033] 1) Plasmid pXYL1-2μ(LEU2)

[0034] Plasmid pXYL1-2μ(LEU2) is to insert xyl1 gene (http: / / www.ncbi.nlm.nih.gov / nuccore / X59465.1) into vector plasmamid-2μ(LEU2) (vector plasmamid-2μ(LEU2) is to insert The pYES2 vector was obtained by replacing the ura selection marker with a leu selection marker, and replacing the GAL1 promoter with a GPD promoter. The pYES2 vector was purchased from Invitrogen Company Catalog no.V825-20, and the sequence of the vector plasmamid-2μ (LEU2) is shown in Sequence Table 1). The vector obtained between the BamH Ⅰ restriction sites.

[0035] The specific construction method is as follows: clone the xylose reductase gene (xyl1) from Pichia stipitis DNA, add BamH Ⅰ sites at both ends, digest the vector plasmamid-2μ (LEU2) with BamH Ⅰ, and connect it to the gene xyl1, Plasmid...

Embodiment 2

[0065] Embodiment 2, the application of Saccharomyces cerevisiae (Saccharomyces cerevisiae) W32N55 CGMCC NO.6090 in the production of ethanol

[0066] 1. Saccharomyces cerevisiae W32N55 CGMCC NO.6090 fermenting xylose to produce ethanol

[0067] 1. Obtaining of seed culture medium

[0068] Inoculate Saccharomyces cerevisiae (Saccharomyces cerevisiae) W32N55CGMCC NO.6090 in 5ml YPD medium, culture overnight at 30°C, 250rpm until saturated, and insert 2% inoculum into 100ml YPD medium with 250ml shake flask (yeast powder 10g / L, peptone 20g / L, glucose 20g / L), 30°C, 250rpm (rotation radius 3cm) for 18h, as the seed culture solution.

[0069] 2. Fermentation

[0070] The seed culture solution obtained in the above 1 was centrifuged at 4500rpm for 7min with a centrifugal radius of 3cm. The above-mentioned seed culture solution was used to collect the bacteria, and the bacteria were resuspended in 100ml of YPX medium (yeast Powder 10g / L, peptone 20g / L, xylose 45g / L), ferment unde...

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Abstract

The invention discloses a Saccharomyces cerevisiae engineering bacterium and its application in the production of ethanol. The invention provides Saccharomyces cerevisiae W32N55, and the preservation number of the Saccharomyces cerevisiae W32N55 is CGMCC NO.6090. The invention also provides an application of the Saccharomyces cerevisiae W32N55 in the preparation of ethanol. Experiments of the invention prove that the engineering bacterium W32N55 provided in the invention is obtained through obtaining the Saccharomyces cerevisiae engineering bacterium by the introducing xylose reductase gene (XYL1) and xylitol dehydrogenase gene (XYL2) to an original strain and by improving the activity of xylulokinase (XK), and through allowing the engineering bacterium containing the XYL1, the XYL2 and XK gene to undergo adaptive acclimation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a brewer's yeast engineering bacterium and its application in ethanol production. Background technique [0002] With the depletion of current energy sources, rising oil prices and global warming caused by the greenhouse effect, people are forced to pay more and more attention to the development and utilization of sustainable energy. At present, lignin resources have become the most abundant renewable resources in the world. If lignin resources can be fully utilized and converted into clean energy, it will definitely become the most beneficial substitute for petroleum. [0003] As a clean and renewable transportation fuel, biofuel based on fuel ethanol is currently one of the most feasible and widely used biological transportation fuels, and it is also one of the most effective ways to reduce fossil fuel consumption and greenhouse gas emissions . Cellulose fuel ethanol technology do...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/18C12P7/06C12P7/10C12R1/865
CPCY02E50/17Y02E50/16Y02E50/10
Inventor 张延平张博张海峰孙红兵李寅马延和
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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