Transgenic engineering saccharomyces cerevisiae SF4 for efficiently fermenting ethanol using xylose

A technology of transgenic engineering and Saccharomyces cerevisiae, which is applied in the field of genetically engineered Saccharomyces cerevisiae SF4, can solve the problems of low ethanol yield, low tolerance to high-concentration sugar, low tolerance to ethanol and fermentation inhibitors, and improve utilization rate effect

Active Publication Date: 2017-05-24
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Although some progress has been made, the following problems still exist in the related transgenic yeast: 1. The yield of ethanol is low; 2. The tolerance of ethanol and fermentation inhibitors is not high; 3. The stability of the recombinant strain is not good; 4. High High sugar tolerance is not high

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  • Transgenic engineering saccharomyces cerevisiae SF4 for efficiently fermenting ethanol using xylose
  • Transgenic engineering saccharomyces cerevisiae SF4 for efficiently fermenting ethanol using xylose
  • Transgenic engineering saccharomyces cerevisiae SF4 for efficiently fermenting ethanol using xylose

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Embodiment Construction

[0060] Below in conjunction with accompanying drawing and embodiment describe in detail:

[0061]1. Application steps of Saccharomyces cerevisiae SF4

[0062] SF4→glycerol tube preservation of strains→YPD medium (2% glucose, 1% yeast extract, 2% peptone, 0.03 g adenine, 2% agar for solids) slope activation→YPD medium liquid triangular flask seeds→according to 0.1 Add ~0.5% of the amount into the straw hydrolyzate or xylose solution → static fermentation for 48-72 hours → distillation of the fermented liquid at 80-100°C → cooling to obtain ethanol.

[0063] 2. Methods and technical routes for the construction and verification of recombinant strains

[0064] The sequence listing shows that SF4 contains xyl1-(G 4 S 1 ) 3 - Nucleotide sequence of xyl2 gene (SEQ ID NO: 5).

[0065] 1. Construction of SF4 and verification of its functions

[0066] 1) pYPGE15XYL1-(G 4 S 1 ) 1 -XYL2 vector construction

[0067] Since the construction of this fusion protein needs to add a seg...

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Abstract

The invention discloses transgenic engineering saccharomyces cerevisiae SF4 for efficiently fermenting ethanol using xylose and relates to the technical field of saccharomyces cerevisiae. The saccharomyces cerevisiae SF4 is a fusion protein gene consisting of two xyloses linked through an oligopeptide chain [-(G4S1)3-] by using enzymes including xylose reductase XYL1 and xylitol oxidase XYL2; the gene sequence of XYL1 is shown in SEQ ID NO:1, the gene sequence of XYL2 is shown in SEQ ID NO:2, and the gene sequence of fusion protein is shown in SEQ ID NO:3. The transgenic engineering saccharomyces cerevisiae SF4 is applied to ethanol fermentation of straw hydrolyzate. The growth amount of the SF4 strain is 2.5 times that of a contrast; during co-fermentation of glucose and xylose, the xylose utilization rate of the SF4 strain reaches 53.8%; during ethanol fermentation of miscanthus sinensis material hydrolyzate, the ethanol producing strain SF4 is increased by about 4.5 times.

Description

technical field [0001] The invention relates to the technical field of Saccharomyces cerevisiae, in particular to a genetically engineered Saccharomyces cerevisiae SF4 that utilizes xylose to efficiently ferment ethanol. Background technique [0002] Xylose is the most abundant five-carbon sugar in straw hydrolyzate, but it cannot be utilized by traditional Saccharomyces cerevisiae. Fully converting xylose into ethanol is a key link in the rational and effective utilization of straw resources. It is theoretically feasible to apply genetic engineering to construct recombinant yeast strains and improve their xylose fermentation ability. Therefore, in the past 30 years, there have been a large number of reports on the application of genetic engineering techniques to improve Saccharomyces cerevisiae at home and abroad. The ability of yeast to ferment xylose. Although some progress has been made, the following problems still exist in the related transgenic yeast: 1. The yield o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81C12N15/62C12R1/865
CPCC12N9/0006C12P7/06C12P7/10C12P2201/00C12Y101/01009C12Y101/01307Y02E50/10
Inventor 彭良才郝勃夏涛涂媛媛王艳婷涂芬熊科魏小洋
Owner HUAZHONG AGRI UNIV
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