Response surface methodology of nitrilase production by recombinant Escherichia coli

• Main Authors: Sachin Dubey, Amit Singh, Uttam C Banerjee
• Format: Article
• Language: English
• Published: Sociedade Brasileira de Microbiologia 2011-09-01
• Series: Brazilian Journal of Microbiology
• Subjects: Biocatalysis; Recombinant E. coli; Nitrilase; Response surface methodology; Central composite design
• Online Access: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000300029

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.