title: Response surface methodology of nitrilase production by recombinant Escherichia coli.
abstract: Growth and nitrilase production by recombinant Escherichia coli cells harbouring pET 21 (b) plasmid, for the expression of Pseudomonas putida nitrilase were improved using response surface methodology. Central composite design was used for obtaining ideal concentration of critical medium components which include fructose, tryptone, yeast extract and lactose. The optimal values for the concentration of fructose, tryptone, yeast extract and lactose were found to be 1.13, 2.26, 3.25 and 0.9 % (w/v), respectively. Here, fructose served as carbon source for the growth while lactose was preferably used as inducer for the expression of foreign protein. Yeast extract in the medium was used as a growth promoter while tryptone was added as a major nitrogen source. Using this optimized medium, an experimental growth of 6.67 (OD at 600 nm) and nitrilase activity of 27.13 U/ml was achieved. This approach for medium development led to an enhancement of the growth and enzyme activity by 1.4 and 2.2 times, respectively, as compared to the un-optimized medium.
authors.affiliation: Biocatalysis Laboratory, Department of Pharmaceutical Technology, National Institute of Pharmaceutical Education and Research , Sector 67, S.A.S. Nagar-160 062, Punjab , India ; School of Pharmaceutical Sciences, University of Geneva & University of Lausanne , 30 Quai Ernest Ansermet, 1211 Geneva , Switzerland.
authors.forename: Sachin
authors.initials: S
authors.lastname: Dubey
authors.forename: Amit
authors.initials: A
authors.lastname: Singh
authors.forename: Uttam C
authors.initials: UC
authors.lastname: Banerjee
keywords: Biocatalysis; Central composite design.; Nitrilase; Recombinant E. coli; Response surface methodology
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