Steroselective hydrolysis of DL-beta-acetylthioisobutyramide catalyzed by genetically engineered E. coli immobilized on Celite 580 in a packed bed bioreactor

Shyh Yu Shaw, Yu Jen Chen, Jung Jung Ou, Lewis Ho

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1607-1613
Number of pages7
JournalJournal of the Chinese Chemical Society
Volume54
Issue number6
DOIs
Publication statusPublished - 2007 Jan 1

All Science Journal Classification (ASJC) codes

  • Chemistry(all)

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