催化学报

催化学报 ›› 2008, Vol. 29 ›› Issue (10): 997-1002.

• 研究论文 • 上一篇    下一篇

内酯酶产生菌的筛选及其催化拆分手性内酯的性能

张仙,潘江,许建和   

  1. 华东理工大学生物反应器工程国家重点实验室, 上海 200237
  • 收稿日期:2008-10-25 出版日期:2008-10-25 发布日期:2008-10-25

Screening of Lactonase-Producing Strains and Their Performance in Catalytic Resolution of Chiral Lactones

ZHANG Xian, PAN Jiang, XU Jianhe*   

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China
  • Received:2008-10-25 Online:2008-10-25 Published:2008-10-25

PDF

1271

被引次数

12

摘要： 以γ-丁内酯系列化合物作为唯一碳源,通过富集培养从土壤中分离得到180株产内酯酶的菌株. 经反复筛选,从中挑选出两株内酯水解活性和选择性均较高的菌株. 经鉴定,这两株真菌皆为镰孢霉菌属,分别命名为Fusarium moniliforme ECU2001和Fusarium proliferatum ECU2002. 对这两株菌的产酶特性进行了研究,发现ECU2001的内酯酶属于胞内酶,而ECU2002的内酯酶同时存在于胞内和胞外. 选择戊二醛交联的方法对这两株菌进行了细胞固定化,结果表明, ECU2002固定化细胞的活力、选择性和稳定性都优于ECU2001. ECU2002固定化细胞反应的适宜温度和pH分别为50 ℃和7.0～7.5. 考察了ECU2002固定化细胞的底物专一性,发现底物为α-羟基-γ-丁内酯时,固定化细胞的活性最高,其催化速率是γ-丁内酯的54.3倍. 利用ECU2002固定化细胞催化α-羟基-γ-丁内酯的对映选择性水解,固定化细胞重复使用10批后活力仍保持为初始活力的92.8%, 水解产物经内酯化后得到(R)-α-羟基-γ-丁内酯的光学纯度为92.8%～96.1% ee.

关键词: 内酯酶, 串珠镰孢菌ECU2001, 层出镰孢菌ECU2002, 固定化细胞, &alpha, -羟基-&gamma, -丁内酯

Abstract: By using immobilized cells of Fusarium proliferatum ECU2002, which was new ly isolated from soil samples, biocatalytic resolution of (±)-α-hydroxy-γ -butyrolactone was performed at a concentration of 1 mol/L and repeated for 10 cycles of reaction, retaining 92.8% of initial activity and affording the produc t with 92.8%-96.1% ee.

Key words: lactonase, Fusarium moniliforme ECU2001, Fusarium proliferatum ECU2002, immobilized cell, &alpha, -hydroxy-&gamma, -butyrolactone