Abstract:
:Nitrilases have attracted tremendous attention for the preparation of optically pure carboxylic acids. This article aims to address the production and utilization of a highly enantioselective nitrilase from Pseudomonas putida MTCC 5110 for the hydrolysis of racemic mandelonitrile to (R)-mandelic acid. The nitrilase gene from P. putida was cloned in pET 21b(+) and over-expressed as histidine-tagged protein in Escherichia coli. The histidine-tagged enzyme was purified from crude cell extracts of IPTG-induced cells of E. coli BL21 (DE3). Inducer replacement studies led to the identification of lactose as a suitable and cheap alternative to the costly IPTG. Effects of medium components, various physico-chemical, and process parameters (pH, temperature, aeration, and agitation) for the production of nitrilase by engineered E. coli were optimized and scaled up to a laboratory scale bioreactor (6.6 l). Finally, the recombinant E. coli whole-cells were utilized for the production of (R)-(-)-mandelic acid.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Banerjee A,Dubey S,Kaul P,Barse B,Piotrowski M,Banerjee UCdoi
10.1007/s12033-008-9094-zsubject
Has Abstractpub_date
2009-01-01 00:00:00pages
35-41issue
1eissn
1073-6085issn
1559-0305journal_volume
41pub_type
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