Molecular cloning of enantioselective ester hydrolase from Bacillus pumilus DBRL-191
- 作者：Shafaq Rasool ; Sarojini Johri ; Syed Riyaz-ul-Hassan ; Qurrat-ul-Ain Maqbool ; Vijeshwar Verma ; Surrinder Koul ; Subhash C. Taneja and Ghulam N. Qazi
- 关键词：Phenyl– ; Sepharose ; Drug intermediates ; Kinetic resolution ; Enantioselective hydrolysis
- 刊名：FEMS Microbiology Letters
- 出版年：2005
- 期刊代码：34_03781097
- 类别：imm
- 出版时间：2005
- 卷：249
- 期：1
- 页码：113-120
- 文件大小：244 K
摘要
A gene from Bacillus pumilus expressed under its native promoter was cloned in Escherichia coli. Recombinant B. pumilus esterase (BPE) affects the kinetic resolution of racemic mixtures such as unsubstituted and substituted 1-(phenyl)ethanols (E 
ncedirect.com/scidirimg/entities/223c.gif" alt="not, vert, similar" border=0> 33–103), ethyl 3-hydroxy-3-phenylpropanoate (E ncedirect.com/scidirimg/entities/223c.gif" alt="not, vert, similar" border=0> 45–71), trans-4-fluorophenyl-3-hydroxymethyl-N-methylpiperidine (E ncedirect.com/scidirimg/entities/223c.gif" alt="not, vert, similar" border=0> 10–13) and ethyl 2-hydroxy-4-phenylbutyrate (E ncedirect.com/scidirimg/entities/223c.gif" alt="not, vert, similar" border=0> 7). The enzyme is composed of a 34-amino acid signal peptide and a 181-amino acid mature protein corresponding to a molecular weight of ncedirect.com/scidirimg/entities/223c.gif" alt="not, vert, similar" border=0>19.2 kD and pI ncedirect.com/scidirimg/entities/223c.gif" alt="not, vert, similar" border=0> 9.4. 3-D the structural model of the enzyme built by homology modelling using the atomic coordinates from the crystal structure of B. subtilis lipase (LipA) showed a compact minimal α/β hydrolase fold.