- 作者: 蕭世裕,陳裕仁,歐蓉蓉,何昇龍
- 作者服務機構: Department of Chemistry, National Cheng Kung University, Tainan, Taiwan, R.O.C. ,Institute of Biotechnology, National Cheng Kung University, Tainan, Taiwan, R.O.C.
- 中文摘要:
Pseudomonas putida IFO12996 catalyzes the stereoselective hydrolysis of methyl DL-b-acetylthioisobutyramide (DL-ATIA) to form D-b-acetylthioisobutyric acid (DAT), a key intermediate for synthesis of a series of angiotensin converting enzyme inhibitors. The esterase gene of Pseudomonas putida IFO12996 was cloned and expressed in Escherichia coli which was further immobilized and retained on a packed bed bioreactor filled with Celite 580. The packed bed bioreactor was used to conduct the stereoselective hydrolysis of DL-ATIA and to give DAT with a yield of 34.5%, enantiometric excess value of 97% and enantioselectivity value > 150. The optimal pH and temperature for the reaction were 9.0 and 57 °C ~ 67 °C, respectively. The kinetic constants (Km and Vmax) of immobilized cells were found to be 372.5 mM and 285.7 mmol min-1 (g cell)-1, respectively. The immobilized cells retained over 60% of the initial catalytic activity after 5 batch cycles of production.
This paper presents a simple, practical and economical process of immobilization of genetically engineered E. coli on a novel packed bed bioreactor for production of DAT. - 英文摘要: --
- 中文關鍵字: Whole cell immobilization; Genetically-engineered E. coli; DL-b-Acetylthioisobutyramide (DL-ATIA); D-b-Acetylthioisobutyric acid (DAT); Packed bed bioreactor.
- 英文關鍵字: --