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Optimization of high cell density fermentation process for recombinant nitrilase production in E. coli

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Title Optimization of high cell density fermentation process for recombinant nitrilase production in E. coli
 
Creator SOHONI, SV
NELAPATI, D
SATHE, S
JAVADEKAR-SUBHEDAR, V
GAIKAIWARI, RP
WANGIKAR, PP
 
Subject HETEROLOGOUS PROTEIN-PRODUCTION
HIGH-LEVEL EXPRESSION
ESCHERICHIA-COLI
PLASMID STABILITY
GROWTH-HORMONE
RNA-POLYMERASE
SYSTEMS
STRATEGIES
CULTURES
GENES
Nitrilase
High cell density fermentation
E. coli BL21 (DE3)
 
Description Nitrilases constitute an important class of biocatalysts for chiral synthesis. This work was undertaken with the aim to optimize nitrilase production in a host that is well-studied for protein production. Process parameters were optimized for high cell density fermentation, in batch and fed-batch modes, of Escherichia coli BL21 (DE3) expressing Pseudomonas fluorescens nitrilase with a T7 promoter based expression system. Effects of different substrates, temperature and isopropyl beta-D-1-thiogalactopyranoside (IPTG) induction on nitrilase production were studied. Super optimal broth containing glycerol but without an inducer gave best results in batch mode with 32 degrees C as the optimal temperature. Use of IPTG led to insoluble protein and lower enzyme activity. Optimized fed-batch strategy resulted in significant improvement in specific activity as well as volumetric productivity of the enzyme. On a volumetric basis, the activity improved 40-fold compared to the unoptimized batch process. (C) 2015 Elsevier Ltd. All rights reserved.
 
Publisher ELSEVIER SCI LTD
 
Date 2016-01-14T13:07:22Z
2016-01-14T13:07:22Z
2015
 
Type Article
 
Identifier BIORESOURCE TECHNOLOGY, 188,202-208
0960-8524
1873-2976
http://dx.doi.org/10.1016/j.biortech.2015.02.038
http://dspace.library.iitb.ac.in/jspui/handle/100/17574
 
Language en