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Yeast mutant and substance production method using the same

一种生产方法、突变体的技术，应用在植物学设备和方法、生物化学设备和方法、发酵等方向，能够解决发酵速度下降、乙醇降低不充分等问题，达到低成本的效果

Active Publication Date: 2011-04-13
TOYOTA JIDOSHA KK
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] On the other hand, there is also a report on improving the yield of the target product by limiting the blocking of the ethanol production pathway to a part and enhancing the metabolic pathway of the target product (Non-Patent Document 5: Saitoh, S (2005) Appl.Environ. Microbiol.71, p2789-2792), however, although the yield of the target product has been improved, there is a problem that the reduction of ethanol is not sufficient, and the fermentation rate is also slightly reduced.

Method used

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  • Yeast mutant and substance production method using the same
  • Yeast mutant and substance production method using the same
  • Yeast mutant and substance production method using the same

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Embodiment 1

[0045] Preparation of DNA fragments for LDH gene introduction / PDC1 gene disruption

[0046] First, a lactate dehydrogenase (LDH) gene was introduced as a foreign gene into yeast as a host, and a DNA fragment for disrupting the pyruvate synthase gene (PDC1 gene) involved in alcohol synthesis was produced.

[0047] Specifically, using the plasmid pBTrp-PDC1-LDHKCB disclosed in JP-A-2003-259878 as a template, the DNA fragment fused with the PDC1 promoter, LDH gene and TDH3 terminator was amplified by PCR. In this PCR, TB215 (5'-GAAACAGCTATGACCATGATTACG-3': SEQ ID NO: 3) and TB1497 (5'-AAGCCTTAAAACGGGAATTCCCCTAAGAAACCAT-3': SEQ ID NO: 4) were used as primers (see figure 1 ).

[0048] On the other hand, the HIS3 gene was amplified using the plasmid pRS403 (available from ATCC) as a template. In this PCR, TB1421 (5'-ATGGTTTCTTAGGGGAATTCCCGTTTTAAGAGCTT-3': SEQ ID NO: 5) and TB422 (5'-GACCAAGTTAGCTGGTCGAGTTCAAAGAGAAAAAAAAAAG-3': SEQ ID NO: 6) were used as primers.

[0049] In add...

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Abstract

The object aims to largely improve the productivity of a desired product and maintain an excellent growth rate and an excellent fermentation rate in the production of the desired product by using yeast. A foreign gene which encodes an enzyme involved in the production of the desired product and HAP4 gene which can be expressed constitutively or a homologous gene thereof are introduced into a yeast host. The yeast mutant is preferably a mutant strain having reduced alcohol productivity compared to its wild-type one.

Description

technical field [0001] The present invention relates to a mutant yeast modified to express a predetermined gene constitutively in a wild-type yeast, and a method for producing a substance using the mutant yeast. Background technique [0002] When using yeast or the like to produce the product of interest, a mutant yeast into which a gene related to the biosynthesis of the product of interest can be introduced in a manner capable of constitutive expression is produced, and the mutant yeast is cultured under appropriate culture conditions, and the yeast is obtained from the cell or the cell. In vitro recovery of target products. In addition, when substances other than ethanol are used as target products, it is preferable to reduce ethanol that is produced in large quantities. So far, in order to reduce ethanol production capacity, strains have been produced that disrupt genes such as pyruvate decarboxylase and alcohol dehydrogenase present in the ethanol production pathway. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/00C12N1/19C12N15/09C12P1/02C12P7/56
CPCC12P7/40C12N9/0006C07K14/395C12P7/04C12N9/88C12P7/56
Inventor 大西彻多田宣纪松下响保谷典子石田亘广嶋村隆
Owner TOYOTA JIDOSHA KK
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