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

Enhancing nitrilase production from Fusarium proliferatum using response surface methodology

“…multivariate designs, allows multiple control variables, is faster to implement and cost effective as compared to traditional univariate approach. As far as nitrilases are concerned, enhancement of nitrilase production using RSM has been studied with only bacterial source (Dubey et al 2011; Shen et al 2009) and fungi have not been studied in this context.…”

“…RSM is a collection of mathematical and statistical method which helps in studying the effects of various parameters at a single time, is faster to implement, time saving and cost effective (Montgomery 2005; Layh et al 1977). Various studies have recently been reported on optimization of nitrilase production in bacterial strains using RSM (Dubey et al 2011; Shen et al 2009). However, to the best of our knowledge, optimization of nitrilase production from fungal strains using RSM has not been reported.…”

Enhanced Production of Nitrilase from Streptomyces sp. MTCC 7546 by Response Surface Method

“…The maximum nitrilase yield Literature survey shows that yeast extract has also been used for nitrilase production from other sources however, the concentration of yeast extract reported in previous works is high (5-10 g/L) [20,21]. Dubey et al [18] used tryptone along with the yeast extract to maximize the production of nitrilase [18]. Medium with high concentration of yeast extract or supplemented with other nitrogen sources may not be economical during large scale production of nitrilase in industries.…”

“…Response surface methodology (RSM) is a statistical method used intensively to study the interaction among variables and optimize the concentration of each component of the medium. It can study the effect of many variables together influencing the response in fewer experiments at lesser time and cost [16][17][18][19]. A few studies are reported on the enhancement of nitrilase production using RSM however, the studies were done on mesophillic sources [20,21].…”

Extracellular L-Asparaginase from Streptomyces labedae VSM-6: Isolation, Production and Optimization of Culture Conditions Using RSM

“…16 Hence statistically engineered optimization of the fermentation process is far more beneficial and admirable to classical methods due to the fact that it scrutinize the interactive influence of the process variables in which a single variable is varied at any time. 17,18 Statistically designed minimum number of the replicates executes the information of the effects of main factors and the viable interactions amid them. 19 Statistical optimization to amplify the enzyme production is persuaded by a number of process variables that are assorted in to two i) media constituents such as carbon, nitrogen minerals and salts ii) cultural parameters such as pH, temperature and incubation time.…”