| Project/Area Number |
08556013
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (A)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
応用微生物学・応用生物化学
|
|---|
| Research Institution | KYOTO UNIVERSITY |
|---|
Principal Investigator |
SHIMIZU Sakayu Kyoto Univ.Agr.Professor, 農学研究科, 教授 (70093250)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
OGAWA Jun Kyoto Univ.Agr.Assistant Prof., 農学研究科, 助手 (70281102)
KATAOKA Michihiko Kyoto Univ.Agr.Assistant Prof., 農学研究科, 助手 (90252494)
KOBAYASHI Michihiko Kyoto Univ.Agr.Senior Lecturer, 農学研究科, 講師 (70221976)
SAKAMOTO Keiji Fuji Chemical Industries.Researcher, 研究所, 主任研究員
|
|---|
| Project Period (FY) |
1996 – 1997
|
| Project Status |
Completed (Fiscal Year 1997)
|
| Budget Amount *help |
¥10,000,000 (Direct Cost: ¥10,000,000)
Fiscal Year 1997: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1996: ¥7,100,000 (Direct Cost: ¥7,100,000)
|
|---|
| Keywords | Lactonase / Fusarium oxysporum / Pantolactone / Enzymatic resolution / Immobilized enzyme |
|---|
| Research Abstract |
A novel enzymatic process for the optical resolution of racemic pantolactone through the stereospecific hydrolysis of D-pantolactone by lactonohydrolase of Fusarium oxysporum is developed. F.oxysporum showed high productivity of the enzyme and the cells containing the enzyme could be used repeatedly for this hydrolysis reaction. D-Pantolactone in a racemic mixture of pantolactone (700mg/ml) was almost stoichiometrically hydrolyzed to D-pantoic acid on incubation with F.oxysporum cells for 24h under conditions in which the pH was automatically maintained at 7.0 (optical purity for D-pantoic acid, 96% e.e.). In practice the hydrolysis of the D-pantolactone in a racemic mixture is achieved by immobilized cells of F.oxysporum as the catalyst. Stable catalysts with high hydrolytic activity can be prepared by entrapping the fungal cells into calcium alginate gels. When the immobilized cells were incubated in a reaction mixture containing 350g/l DL-pantolactone for 21h at 30ﾟC under the conditions of automatic pH control (pH 6.8-7.2), 90-95% of the D-pantolactone was hydrolyzed (optical purity, 90-97% e.e.). After repeated reactions for 180 times (i.e., 180 days), the immobilized mycelia retained more than 90% of their initial activity. The enzymatic resolution process of racemic pantolactone can skip several tedious steps which are necessary in the conventional chemical resolution process, and this is a considerable practical advantage.
|
