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Recombinant yeast and branched alcohol production method using recombinant yeast

a technology of branched alcohol and recombinant yeast, which is applied in the direction of microorganisms, biochemical equipment and processes, enzymes, etc., can solve the problems of poor oxidative stability, high cost, and a shortage of amino acids necessary for microorganism growth, and achieve the effect of low cos

Inactive Publication Date: 2010-12-09
TOYOTA JIDOSHA KK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0012]The recombinant yeast of the present invention has an excellent feature of producing a desired branched alcohol that can be used as an automotive fuel and a resin material. With the use of the recombinant yeast of the present invention, the above branched alcohol can be produced at low cost.

Problems solved by technology

However, regarding branched alcohol production with the use of the amino acid synthesis pathway, the supply shortage of amino acids necessary for microorganism growth is a matter of concern.
There are still problems for mass-production of branched alcohol, which is a product of interest.
However, the resulting branched alcohol has unsaturated bonds, and therefore the oxidative stability is poor when such alcohol is used as fuel, which is problematic.
However, such step is cost-consuming, which is also problematic.

Method used

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  • Recombinant yeast and branched alcohol production method using recombinant yeast
  • Recombinant yeast and branched alcohol production method using recombinant yeast
  • Recombinant yeast and branched alcohol production method using recombinant yeast

Examples

Experimental program
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Effect test

example 1

Production of the HMG1 / YPH499 Strain

[0046]In this Example, Saccharomyces cerevisiae capable of causing high expression of the HMG-CoA reductase gene was produced first. Specifically, the HMG-CoA reductase gene derived from the Saccharomyces cerevisiae DBY746 strain was introduced into the Saccharomyces cerevisiae YPH499 strain. A detailed explanation is as follows.

(1) Preparation of pRS404Tcyc and pRS406Tcyc

[0047]A CYC1 transcription terminator CYC1t fragment was prepared by PCR with the use of a pair of primers designed as shown below.

XhoI-Tcyc1FW:(SEQ ID NO: 5)5′ TGC ATC TCG AGG GCC GCA TCA TGT AAT TAG 3′ApaI-Tcyc1RV:(SEQ ID NO: 6)5′ CAT TAG GGC CCG GCC GCA AAT TAA AGC CTT CG 3′

[0048]In addition, pYES2 (Invitrogen) (0.1 mg) as a PCR template and the above pair of primers (50 pmol each) were used. The PCR reaction solution was a solution (50 ml) containing 1×pfu buffer with MgSO4 (Promega), 10 nmol dNTP, a Pfu DNA polymerase (Promega) (1.5 u), and a perfect match polymerase enhance...

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Abstract

This invention provides a recombinant yeast that can produce branched alcohol appropriate for automotive fuel and the like and a branched alcohol production method whereby branched alcohol can be produced at low cost with the use of the recombinant yeast.A recombinant yeast in which a hydroxymethyl glutaryl-CoA reductase gene has been expressed to a high degree and the ADP-ribose pyrophosphatase gene and / or the yhfR gene are introduced so as to be expressed therein is provided.

Description

BACKGROUND OF THE INVENTION[0001]1. Field of the Invention[0002]The present invention relates to a recombinant yeast having a predetermined gene introduced therein and a branched alcohol production method using the recombinant yeast.[0003]2. Background Art[0004]In recent years, the following problems have been arising regarding ethanol used as automotive fuel: the energy density thereof is low and the characteristics thereof significantly differ from those of conventional petroleum fuel. Therefore, studies have been conducted to produce butanol or the like, which is an alcohol with an extended carbon chain length serving as fuel characteristically closer to petroleum fuel, by a fermentative production method with the use of Escherichia coli and yeast (see WO 2007 / 041269 and WO 2008 / 052991). In addition, since a gasoline fuel with the high octane number of such a straight-chain alcohol is preferably used, fuels comprising 2-methyl-1-butanol, 3-methyl-1-butanol, and the like have been...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12P7/04C12N1/19
CPCC12N9/0006C12P7/04C12N15/52C12N9/1241
Inventor YONEDA, SATOSHIMURAMATSU, MASAYOSHIOBATA, SHUSEIINOUE, AI
Owner TOYOTA JIDOSHA KK
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