Optimization of Thermo-Alkali Stable Amylase Production and Biomass Yield from Bacillus sp. Under Submerged Cultivation

Ameer Khusro,Chirom Aarti,Paul Agastian,Kaliyan Barathikannan

doi:10.3390/fermentation3010007

Abstract

The present context was investigated to optimize amylase production and cell biomass of poultry-associated Bacillus sp. using a conventional as well 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 (R2) 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 from Bacillus 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.

Highlights

Amylases constitute a group of enzymes that yields dextrin and numerous monomer products by hydrolysing α-1,4 glycosidic linkages of starch molecules
Isolation and Identification of Amylase-Producing Bacteria hyperamylase-producing bacterium was isolated from the feces soil sample of poultry farm based on 3.1
The bacterial morphological and biochemical characteristics the isolate based on assay the standard colony confirmed the production of amylase by of further qualitative enzyme using the Bergey’s streak plate technique (Figure 1a)

Summary

Introduction

Amylases constitute a group of enzymes that yields dextrin and numerous monomer products by hydrolysing α-1,4 glycosidic linkages of starch molecules. These amylolytic enzymes are of great significance in many biotechnological processes including pharmaceutical, food, textile, fibre, detergent, brewing, oil drilling and paper industries [1]. Amylase (EC 3.2.1.1) production from microbial sources have gained immense interest among researchers because of bulk production capability and the easy manipulation of the microorganisms in order to produce enzymes with the desired characteristics [2]. Is an industrial microorganism that has been exploited commercially due to short fermentation cycle, consistency, safe handling, eco-friendly characteristics, easy manipulation, enhanced enzymatic activity at harsh conditions, cost-effective enzyme production and ability to secrete secondary Bacillus sp. is an industrial microorganism that has been exploited commercially due to short fermentation cycle, consistency, safe handling, eco-friendly characteristics, easy manipulation, enhanced enzymatic activity at harsh conditions, cost-effective enzyme production and ability to secrete secondary

Methods

Results

Discussion

Conclusion