1994 Fiscal Year Final Research Report Summary
New Stereospecific Synthesis of Bioactive Compounds Using Lipase as Catalyst in Organic Solvent
| Project/Area Number |
04556014
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
製造化学・食品
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
ODA Junichi Institute for Chemical Research, Kyoto University, Professor, 化学研究所, 教授 (50027041)
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| Co-Investigator(Kenkyū-buntansha) |
HIRATAKE Jun Inst.Chem.Res., Kyoto Univ., Research Associate, 化学研究所, 助手 (80199075)
KATO Hiroaki Inst.Chem.Res., Kyoto Univ., Research Associate, 化学研究所, 助手 (90204487)
NISHIOKA Takaaki Inst.Chem.Res., Kyoto Univ., Associate Professor, 化学研究所, 助教授 (80026559)
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| Project Period (FY) |
1992 – 1994
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| Keywords | lipase / stereospecific synthesis / organic solvent / cyanohydrin / transesterification / lipase activator / 活性化因子 / 大量発現 |
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| Research Abstract |
Using lipases as catalyst in organic solvent, optically pure cyanohydrins in (s) -configuration were stereoselectively synthesized by transesterification reaction in almost 100% chemical yeild. When disaccharide such as trehalose or sucrose was added at the immobilization of lipases, the carbohydrate added keeps lipase in optimal hydration state and increases the maximal lipase activity. The addition of carbohydrate also increases the durability of the enzyme in organic solvent. Lipase was inhibited by organophosphorus esterase inhibitors in organic solvent, but not inhibited in aqueous solvent. This indicates that the lipase catalyzes the transesterification reaction as a typical serine esterase in organic solvent. Lipase and its activator genes from Pseudomonas aeruginosa TE3285 were cloned and expressed in Escherichia coli. The product lipase was not secreted, but stored in the cells as inactive inclusion bodies. The activator protein was expressed and purified. The lipase activity were recovered by refolding the inclusion bodies under the presence of the activator protein. They are reactivated through a 1 : 1 complex without ATP.
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