Response surface methodology: An effective optimization strategy for enhanced production of nitrile hydratase (NHase) by Rhodococcus rhodochrous (RS-6)

Abstract

Nitrile hydratase is an enzyme which catalyze the hydration of nitriles into amide and their role as catalysts for acrylamide production in industries are well known. The present study aims at statistically optimizing physiological and nutritional parameters for NHase production from Rhodococcus rhodochrous (RS-6). The effect of incubation period, temperature, pH, carbon and nitrogen sources on the production of NHase was investigated by one factor at a time strategy. Further optimization process was carried out by response surface methodology for studying the interactive effect of these variables using central composite design. The optimized levels of variables obtained by statistical analysis were: incubation period 48 h, temperature 33 °C, pH 7.0, glycerol 1% and urea 0.75%, which resulted in maximum NHase production. The results of ANOVA were significant with the F-value of the model being 296.78, value of R2 is 0.9983 and the lack of fit test was not significant. The contour and response surface plots showed significant interaction between the variables. The NHase yield was enhanced up to 6.22 fold by statistical optimization using RSM. Thus, the developed experimental design is effective towards process optimization for NHase production from R. rhodochrous (RS-6).