Production method for ethanol using recombined yeast

A manufacturing method and technology of recombinant yeast, applied in the direction of recombinant DNA technology, fermentation, fungi, etc., can solve the problems of unreported and unreported acetic acid assimilation, achieve excellent ethanol yield, reduce the amount of entrainment, and maintain ethanol yield Effect

Inactive Publication Date: 2015-11-18
TOYOTA JIDOSHA KK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few reports in these literatures on the assimilation of acetic acid during the assimilation of xylose
[0018] As mentioned above, there is no report in the prior art that efficiently metabolizes and decomposes acetic acid under the condition of ethanol fermentation while assimilating xylose

Method used

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  • Production method for ethanol using recombined yeast
  • Production method for ethanol using recombined yeast
  • Production method for ethanol using recombined yeast

Examples

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Embodiment 1

[0142] In this example, a recombinant yeast into which a xylose isomerase gene and an Escherichia coli acetaldehyde dehydrogenase gene (mhpF gene) were introduced was produced, and the acetate metabolism ability of the recombinant yeast was evaluated.

[0143]

[0144] (1) Vector for XKS1 gene introduction

[0145] As a vector for yeast introduction of xylulokinase (XK) gene derived from Saccharomyces cerevisiae (S.cerevisiae), figure 1 pUC-HIS3U-P_HOR7-XKS1-T_TDH3-P_TDH2-hph-T_CYC1-HIS3D indicated. This vector contains: the XKS1 gene (genebank: X61377) which is an XK gene derived from Saccharomyces cerevisiae (S. TDH2 promoter and 3' Hygromycin phosphotransferase (hph) gene (marker gene) flanked by CYC1 terminator. In addition, a restriction enzyme Sse8387I site was introduced outside the homologous recombination region. In addition, the nucleotide sequence of the coding region of the XKS1 gene derived from S. cerevisiae strain NBRC304 and the amino acid sequence of xyl...

Embodiment 2

[0184] In this example, the xylose isomerase gene and the mhpF gene and adhE gene of Escherichia coli, the acetaldehyde dehydrogenase gene derived from Clostridium beijerinckii (Clostridium beijerinckii) or the gene derived from Chlamydomonas reinhardtii were produced. Recombinant yeast with aldehyde dehydrogenase gene. In the recombinant yeast produced in this example, one or both of the intrinsic pair of ADH2 genes were disrupted.

[0185]

[0186] (1) Plasmids for XI·XKS1·TKL1·TAL1·RKI1·RPE1 gene introduction and GRE3 gene disruption

[0187] The GRE3 gene was destroyed at the GRE3 locus, and at the same time, the amino acid at position 377 of the xylose isomerase gene derived from the intestinal protist of Reticulitermessperatus was replaced from asparagine to cysteine. Mutant gene with increased sugar assimilation rate (XI_N337C), yeast-derived xylulokinase (XKS1) gene, pentose phosphate pathway transketolase 1 (TKL1) gene, transaldolase 1 (TAL1) gene , ribulose phospha...

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Abstract

　In order to utilize xylose in yeast having a xylose-metabolizing capacity and reduce the concentration of acetic acid by metabolizing acetic acid in the medium during ethanol fermentation, this production method for ethanol comprises a step for performing ethanol fermentation by culturing, in a medium containing xylose, recombined yeast into which a xylose isomerase gene and an acetaldehyde dehydrogenase gene have been introduced.

Description

technical field [0001] The present invention relates to a method for producing ethanol using a recombinant yeast capable of metabolizing xylose. Background technique [0002] Cellulosic biomass is effectively utilized as a raw material for useful alcohols such as ethanol and / or organic acids. In order to increase the yield of ethanol in the production of ethanol using cellulosic biomass, yeasts capable of utilizing the 5-carbon monosaccharide xylose as a substrate have been developed. For example, Patent Document 1 discloses that a xylose reductase gene and a xylitol dehydrogenase gene derived from Pichiastipitis and a xylulokinase gene derived from S. cerevisiae are inserted into the chromosome of yeast. [0003] In addition, it is known that acetic acid is contained in a large amount in the hydrolyzate of cellulosic biomass and suppresses ethanol fermentation by yeast. In particular, it is known that ethanol fermentation using xylose as a sugar source is significantly s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N1/19C12P7/06
CPCC12N1/16C12N9/0008C12N9/90C12P7/10Y02E50/17Y02E50/10
Inventor 大西彻多田宣纪保谷典子片平悟史石田亘广名仓理纱
Owner TOYOTA JIDOSHA KK
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