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Highly productive isopropyl alcohol-producing bacterium

A technology of isopropanol and isopropanol dehydrogenase, applied in the field of isopropanol-producing bacteria, can solve the problem that glucose cannot be completely inhibited

Active Publication Date: 2012-11-21
MITSUI CHEM INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, since the above-mentioned compounds are sometimes necessary for the growth and reproduction of Escherichia coli, the amount of glucose consumed by the above-mentioned side reactions cannot be completely suppressed.

Method used

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  • Highly productive isopropyl alcohol-producing bacterium
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  • Highly productive isopropyl alcohol-producing bacterium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0119]

[0120] The pntA gene promoter on the genome of the Escherichia coli B strain was replaced with the GAPDH promoter to enhance the expression of the pntA gene.

[0121] The full base sequence of the genomic DNA of Escherichia coli MG1655 strain is known (GenBank Accession No. U00096), encoding NAD(P) of Escherichia coli + The base sequence of the gene for the α subunit of transhydrogenase (AB-specific) (hereinafter, sometimes abbreviated as pntA) has also been reported (GenBank Accession No. X04195). It is also known that pntA forms membrane transhydrogenase β subunit (pntB) and an operon on the genomic DNA of Escherichia coli MG1655 strain.

[0122]As the nucleotide sequence of the promoter necessary for the expression of the above-mentioned gene, the glyceraldehyde 3-phosphate dehydrogenase derived from Escherichia coli described in 397-440 in the nucleotide sequence information of GenBank Accession No. X02662 can be used (hereinafter, sometimes referred to as GAPD...

Embodiment 2

[0131]

[0132] As described below, making fortified NAD(P) + Expression of transhydrogenase (AB-specific) gene (pnt) in isopropanol-producing E. coli.

[0133] pGAP-Iaaa / B::pnt strain was obtained by transforming pGAP-Iaaa described in Example 4 of WO2009 / 008377 into the B::pnt strain prepared in Example 1. pGAP-Iaaa is an expression vector plasmid having the function of converting the E. coli-derived thiolase gene, E. coli-derived CoA transferase gene, Clostridium acetobutylicum-derived acetoacetate decarboxylase gene, and Clostridium beijerinckii Source The isopropanol dehydrogenase gene was strongly expressed using a promoter of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) derived from Escherichia coli. The production method of pGAP-Iaaa is clearly described in Example 4 of WO2009 / 008377.

Embodiment 3

[0135]

[0136] CoA transferase genes (atoD and atoA) and thiolase genes (atoB) derived from Escherichia coli form an operon on the genome of Escherichia coli in the order of atoD, atoA, atoB (Journal of Baceteriology Vol.169 pp 42-52 Lauren Sallus Jenkins et al.), therefore, by changing the promoter of atoD, the expression of CoA transferase gene and thiolase gene can be controlled simultaneously. Therefore, Escherichia coli in which the promoter of the atoD gene on the genome of the host Escherichia coli was replaced with the GAPDH promoter and the expressions of the atoD, atoA, and atoB genes on the genome were enhanced was produced.

[0137] The complete base sequence of the genomic DNA of the Escherichia coli MG1655 strain is known (Genbank Accession No. U00096), and the base sequence of the gene encoding the α subunit of CoA transferase of the Escherichia coli MG1655 strain (hereinafter, sometimes abbreviated as atoD) There are also reports. That is, atoD is described...

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Abstract

The invention provides an Escherichia coli which comprises an isopropyl alcohol production system. The isopropyl alcohol-producing Escherichia coli has at least one enhanced enzymatic activity that is selected from the group consisting of enhanced malate dehydrogenase activity, enhanced NAD(P)+ transhydrogenase (AB-specific) activity and enhanced thiolase activity. Also disclosed is a method for producing isopropyl alcohol, which comprises a process wherein isopropyl alcohol is produced from a plant-derived starting material using the above-described isopropyl alcohol-producing Escherichia coli.

Description

technical field [0001] The present invention relates to an isopropanol-producing bacterium and a method for producing isopropanol using the same. Background technique [0002] Propylene is an important basic raw material for synthetic resins such as polypropylene and petrochemicals, and is widely used in automotive shock absorbers, food containers, films, and medical devices. [0003] Since isopropanol produced from plant-derived raw materials can be converted to propylene through a dehydration process, it is promising as a carbon-neutral propylene raw material. According to the Kyoto Protocol, all developed countries are obliged to reduce carbon dioxide emissions by 5% compared with 1990 between 2008 and 2012. In this case, carbon-neutral propylene is extremely important to the global environment due to its wide applicability . [0004] Bacteria that assimilate plant-derived materials to produce isopropanol are known. For example, International Publication No. 2009 / 00837...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12P7/04C12N15/09
CPCC12N9/1029C12P7/04C12N9/0036C12N15/70C12N9/0006
Inventor 松本佳子平野淳一郎森重敬白井智量高桥均天野仰竹林望和田光史清水浩古泽力平泽敬
Owner MITSUI CHEM INC
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