Development of enzymatic resolution process of pantolactone by fungal lactonase

真菌内酯酶分解泛内酯工艺的研究进展

基本信息

批准号：
08556013
负责人：
SHIMIZU Sakayu
金额：
$ 6.4万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08556013/
关键词：
Lactonase Fusarium oxysporum Pantolactone Enzymatic resolution Immobilized enzyme

项目摘要

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.

开发了一种新型的酶促方法，通过通过牛孢子的乳糖水解型牛乳糖酶进行D-Pantolactone的立体水解，用于消除外星型摇托酮的光学过程。 F. oxysporum显示出酶的高生产率，并且含有该酶的细胞可反复用于这种水解反应。 D-Pantolactone中的D-Pantolactone（700mg/ml）几乎将其水解为D-Pantoic酸在与F.Oxysporum细胞一起孵育24H时，在24H的条件下自动维持为24H（在7.0处保持pH值为7.0（可用于D-Pantoic Acid，D-Pantoic Acid），96％E.e.e。在实践中，通过固定的f. oxysporum作为催化剂的细胞来实现DASEMIC混合物中D-苯甲酸的水解。具有高水解活性的稳定催化剂可以通过将真菌细胞置于藻酸钙凝胶中来制备。当在自动pH对照条件下，将固定的细胞在含有350g/l dl-pantolactone的反应混合物中孵育30°C（pH 6.8-7.2）时，将90-95％的D-pantolactone孵育为30°C（pH 6.8-7.2），将D-Pantolactone水解为水解（光学纯度，90-97％E.E.）。在重复反应180次（即180天）后，固定的菌丝体保留了其初始活性的90％以上。外消旋胸肌的酶促分辨率过程可以跳过几个乏味的步骤，这些步骤在常规化学分辨率过程中是必需的，这是一个可观的实际优势。