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Steroselective Hydrolysis of DL‐Beta‐Acetylthioisobutyramide Catalyzed by Genetically Engineered E. Coli Immobilized on Celite 580 in a Packed Bed Bioreactor
Author(s) -
Shaw ShyhYu,
Chen YuJen,
Ou JungJung,
Ho Lewis
Publication year - 2007
Publication title -
journal of the chinese chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.329
H-Index - 45
eISSN - 2192-6549
pISSN - 0009-4536
DOI - 10.1002/jccs.200700227
Subject(s) - chemistry , bioreactor , pseudomonas putida , hydrolysis , esterase , packed bed , chromatography , yield (engineering) , catalysis , immobilized enzyme , escherichia coli , stereoselectivity , enzyme , organic chemistry , biochemistry , gene , materials science , metallurgy
Abstract Pseudomonas putida IFO12996 catalyzes the stereoselective hydrolysis of methyl dl ‐β‐acetylthioisobutyramide ( dl ‐ATIA) to form d ‐β‐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 μmol 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.