Enzymatic resolution of methyl DL-β-acetylthioisobutyrate and DL-β-acetylthioisobutyramide using a stereoselective esterase from Pseudomonas putida IFO12996

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

研究成果: Article同行評審

8 引文 斯高帕斯(Scopus)

摘要

Esterase (PpEST) from Pseudomonas putida IFO12996 catalyzes the stereoselective hydrolysis of methyl dl-β-acetylthioisobutyrate (DL-MATI) and dl-β-acetylthioisobutyramide (DL-ATIA) to give d-β- acetylthioisobutyric acid (DAT). DAT is a key intermediate for the synthesis of a series of angiotensin converting enzyme inhibitors. To use enzyme for the DAT production, the PpEST gene of P. putida IFO12996 was cloned and expressed in Escherichia coli. PpEST with a molecular weight of 33 kDa could hydrolyze DL-MATI and DL-ATIA to give DAT with enantiometric excess value (e.e. value) about 97% and enantioselectivity value (E-value) >150, respectively. The kinetic constants of PpEST for DL-MATI and DL-ATIA were examined and they showed that DL-ATIA was a poorer substrate than DL-MATI for PpEST. However, DL-ATIA was 20-fold more soluble in water than DL-MATI, it was more stable than DL-MATI and it did not show substrate inhibition of the PpEST up to 780 mM. This result suggested that PpEST is an esterase but with amidase activity, which can kinetically resolve DL-ATIA to yield DAT and DL-ATIA is a better choice than DL-MATI for industrial production of DAT by the enzymatic resolution method.

原文English
頁(從 - 到)163-170
頁數8
期刊Journal of Molecular Catalysis B: Enzymatic
38
發行號3-6
DOIs
出版狀態Published - 2006 三月 15

All Science Journal Classification (ASJC) codes

  • 催化
  • 生物工程
  • 生物化學
  • 製程化學與技術

指紋

深入研究「Enzymatic resolution of methyl DL-β-acetylthioisobutyrate and DL-β-acetylthioisobutyramide using a stereoselective esterase from Pseudomonas putida IFO12996」主題。共同形成了獨特的指紋。

引用此