| Project/Area Number |
17580070
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | Kyoto Prefectural University |
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Principal Investigator |
MATSUI Hiroshi Kyoto Prefectural University, Graduate School of Agriculture, Professor, 農学研究科, 教授 (40336730)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Kunihiko Kyoto Prefectural University, Graduate School of Agriculture, Associate Professor, 農学研究科, 助教授 (90184001)
TSUJIMOTO Yoshiyuki Kyoto Prefectural University, Graduate School of Agriculture, Assistant Professor, 農学研究科, 助手 (20315930)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Purinenucleotide Biosynthesis / Purinenucleotide Production / Purinenucleoside Production / Bacillus subtilis / Corynebacterium ammoniagenes / E.coli / Glucose-6-P isomerase / Glucose-6-P dehydrogenase / Corynebacterium ammoniagenes / 好熱性細菌 |
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| Research Abstract |
Our final research purpose is to investigate ability to produce purine nucleotide or nucleoside in three kinds of bacteria, Bacillus subtilis, Corynebacterium ammoniagenes, and Escherichia coli. Thus, we have carried out biochemical analysis of metabolic flux in glycolysis and pentose phosphate cycles, glucose uptake system, and excretion mechanism of purine nucleotide. The results obtained until now are described below.
1. We obtained the enzymatic parameter data of glucose-6-phosphate isomerase and glucose-6-phosphate dehydrogenase related to the distribution of glucose-6-phosphate, in order to investigate the properties of metabolic flux in glycolysis and pentose phosphate cycles into purine nucleotide biosynthesis. Furthermore, we analyzed an intracellular glucose-6-phosphate isomerase and glucose-6-phosphate dehydrogenase, and an intracellular pool of glucose-6-phosphate. From these results, we can see capability of the metabolic flux to purine nucleotide biosynthesis in order of Bacillus subtilis> Corynebactrium glutamicum> E.coli.
2. We investigated inosine-producing ability in E.coli, using the breeding method by gene engineering. It was shown that inosine productivity by improving only purine nucleotide biosynthetic pathway was very low in E.coli. The inosine productivity could be improved by shutdown of glycolysis and Entner-Doudoroff pathways. Furthermore, the existence of over three kinds of inosine-degrading enzymes was found, and it was suggested that these enzymes were an important factor on inosine productivity.
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