Enzymatic preparation of ^<13>C-labelled compounds with C_1-fix ing system of a methylotroph
| Project/Area Number |
04660118
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用微生物学・発酵学
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| Research Institution | Tottori Uniersity |
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Principal Investigator |
KITA Keiko (1993) Tottori University Faculty of Engineering Accoc. Prof., 工学部, 助教授 (70234226)
加藤 暢夫 (1992) 鳥取大学, 工学部, 教授 (50026556)
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| Co-Investigator(Kenkyū-buntansha) |
KATO Nobuo Kyoto University Faculty of Agriculture Prof., 農学部, 教授 (50026556)
YANASE Hideshi Tottori University Faculty of Engineering Accoc. Prof., 工学部, 助教授 (20158033)
喜多 恵子 鳥取大学, 工学部, 教務職員 (70234226)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | ^<13>C-Labelled sugar phosphate / Methanl / Formaldehyde / Methyomonas aminofaciens / C_1-fixing system / Alcohol oxidase / Catalase / 安定同位体標識化合物 / ^<13>C-グルコース / ^<13>C-フルクトース / メチロトローフ / リブロースモノリン酸経路 |
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| Research Abstract |
[^<13>C]Formaldehyde was selectively incorporated into the C-1 position of D-fructose 6-phosphate by condensation with D-ribose 5-phosphate catalyzed by a partially purified enzymesystem for formaldehyde fixation in Methylomonas sminofaciens 77a. Much of the [1-^<13>C]D-fructose 6-phosphate production in this reaction was converted to [1-^<13>C]D-glucose 6-phosphate by the addition of glucose-6-phosphate isomerase. A fed-batch reaction with periodic additions of the substrates afforded 56.2 g/liter D-glucose 6-phosphate and 26.8 g/iter D-fructose 6-phosphate. When [^<13>C]methanol was used as the C_1-donor, the yield of [1-^<13>C]D-glucose 6-phosphate was high when alcohol oxidase was added. The optimum conditions for sugar phosphate production in the fed-batch reaction gave 45.6 g/liter [1-^<13>C]D-glucose 6-phosphate and 16.4 g/liter [1-^<13>C]D-fructose 6-phosphate in 165 min. The catalase to the reaction system supplied molecular oxygen for methanol oxidation to formaldehyde by alcohol oxidase.
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Report
(3 results)
Research Products
(11 results)
