Investigation of Various Genotype Characteristics for Inosine Accumulation in Escherichia coli W3110
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概要
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For the derivation of an inosine-overproducing strain from the wild type microorganism, it is known that the addition of an adenine requirement, removal of purine nucleoside hydrolyzing activity, removal of the feed-back inhibition, and repression of key enzymes in the purine nucleotides biosynthetic pathway are essential. Thus, the disruption of pur A (adenine requirement), deoD (removal of purine nucleosides phosphorylase activity), purR (derepression of the regulation of purine nucleotides biosynthetic pathway), and the insensitivity of the feedback inhibition of phosphoribosylpyrophosphate (PRPP) amidotransferase by adenosine 5'-monophosphate (AMP) and guanosine 5'-monophosphate (GMP) were done in the Escherichia coli strain W3110,and then the inosine productivity was estimated. In the case of using a plasmid harboring the PRPP amidotransferase gene (purF) that encoded a desensitized PRPP amidotransferase, purF disrupted mutants were used as the host strains. It was found that the innovation of the four genotypes brought about a small amount of inosine accumulation. Furthermore, an adenine auxotrophic mutant of E. coli showed inappropriate adenine use because its growth could not respond efficiently to the concentration of adenine added. As the presence of adenosine deaminase is well known in E. coli and it is thought to be involved in adenine use, a mutant disrupted adenosine deaminase gene (add) was constructed and tested. The mutant, which is deficient in purF, purA, deoD, purR, and add genes, and harboring the desensitized purF as a plasmid, accumulated about 1 g of inosine per liter. Although we investigated the effects of purR disruption and purF gene improvement, unexpectedly an increase in the inosine productivity could not be found with this mutant.
- 社団法人日本農芸化学会の論文
- 2001-03-23
著者
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Matsui H
Graduate School Of Science Osaka University
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Minaguchi Hiroshi
Department Of Obstetrics And Gynecology Yokohama City University School Of Medicinedepartment Of Obs
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Matsui Hiroshi
Graduate School Of Science Osaka University
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Kawasaki Haruhiko
Department Of Applied Biochemistry Faculty Of Agriculture Osaka Prefecture University
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Kawasaki Haruhiko
Department Of Applied Biochemistry Faculty Of Agriculture Osaka Prefecture Univeisity
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Kawasaki Haruhiko
Department Of Applied Biochemistry Faculty Of Agriculture University Of Osaka Prefecture
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Kawasaki H
Department Of Environmental Materials Science Tokyo Denki University
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Shimaoka Megumi
Fermentation and Biotechnology Laboratories, Ajinomoto Co., Inc.
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Kurahashi Osamu
Fermentation and Biotechnology Laboratories, Ajinomoto Co., Inc.
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KAWASAKI Hisashi
Fermentation and Biotechnology Laboratories, Ajinomoto Co., Inc.
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MATSUI Hiroshi
Fermentation and Biotechnology Laboratories, Ajinomoto Co., Inc.
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Shimaoka M
Fermentation And Biotechnology Laboratories Ajinomoto Co. Inc.
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Shimaoka Megumi
Fermentation And Biotechnology Laboratories Ajinomoto Co. Inc.
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Kurahashi O
Fermentation And Biotechnology Laboratories Ajinomoto Co. Inc.
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Kurahashi Osamu
Fermentation & Biotechnology Laboratories
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Matsui Hiroshi
Physics Department Graduate School Of Science Tohoku University:center For Interdisciplinary Researc
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Muranaka Hiroshi
Graduate School Of Science Osaka University
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Matsui Hiroshi
Physics Department Graduate School Of Science Tohoku University:center For Interdisciplinary Researc
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