Optimization of thermo-alkali stable amylase production and biomass yield from bacillus sp. under submerged cultivation

Abstract

The present context was investigated to optimize amylase production and cell biomass of poultry-associated Bacillus sp. using a conventional aswell as statistical approach. Box-Behnken design (BBD) matrix at N = 29 was employed to optimize four independent variables, selected from one factor at a time (OFAT) technique, for maximum amylase production and biomass yield. The relative activity of crude amylase obtained from the isolate showed stability at high temperature (60 ° C) and alkaline condition (pH 9) up to 4 h of incubation, thereby indicating its alkali-tolerant and thermo-stable property. The BBD resulted in enhanced amylase activity of 145.32 U/mL when the basal medium was slightly acidic (pH 6) and kept at a temperature of 35 ° C with the shaking speed of 130 rpm, in addition to being incubated for 24 h. The selected factors, when employed with this statistical optimization approach, showed 1.5-fold and 2-fold enhancements in the amylase production and biomass yield respectively compared to the OFAT method. Analysis of variance (ANOVA) revealed high coefficient of determination (R 2 ) of 0.96 to 0.99 for both the responses at significant level (p < 0.05). Three-dimensional response and 2D contour plot of the quadratic model showed interdependent interaction between the effective variables. Long-term thermo-alkali stability of amylase obtained fromBacillus sp. suggested not only its wide applications in pharmaceutical, food and biotechnological industries, but also suggested a potent replacement of existing amylases on the market.