A marked enhancement in production of amylase by Bacillus amyloliquefaciens in flask fermentation using statistical methods

Abstract

A total of 126 bacterial strains were isolated from soil samples. Among them, 11 isolates were found positive for amylase production. Strain YL produced the largest zone of clearance on plate assay. The isolate YL was identified as Bacillus sp. based on morphological and physiochemical characterization. According to 16S rRNA gene sequencing data, the closest phylogenetic neighbor of strain YL was Bacillus amyloliquefaciens (99.54%). After that, an optimization of culture conditions was carried out for the improvement of α-amylase production. Response surface methodology (RSM) was applied to evaluate the effect of medium components including wheat bran, cottonseed extract, yeast extract, starch, NaCl and CaCl 2 . Three variables (wheat bran, cottonseed extract, and starch), which were identified to significantly affect amylase production by Plackett-Burman design were further optimized using response surface methodology of Box-Behnken design (BBD). The optimal concentrations estimated for each variable related to the maximum of amylase activity (86 kU/mL) were 10.80 g/L wheat bran, 9.90 g/L cottonseed extract, 0.5 g/L starch, 2.0 g/L yeast extract, 5.00 g/L NaCl and 2.00 g/L CaCl 2 . The fermentation using optimized culture medium allowed a significant increase in amylase production (by 3-fold). The improvement in the α-amylase production after optimization process can be considered adequate for large-scale applications.

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

“…Hence, there is an urgent need to improve enzyme productivity by optimizing physical factors and medium components required for the growth and utilization of bacteria at the industrial scale. Zhao et al [29] and Prajapati et al [30] reported similar studies on the enhancement in amylase production by Bacillus sp. using statistical tools.…”

Microbial Alpha-Amylase Production: Progress, Challenges and Perspectives

“…57 On the other hand, design expert (version 8.0) is frequently utilized to improve the alpha-amylase production from thermostable and alklophilic alpha-amylase from Bacillus amyloliquefaciens KCP2. 51 Box-Behnken designs: The Box-Behnken designs have been used for RSM experiment to enhance alpha-amylase production from Aspergillus oryzae S2, Aspergillus oryzae CBS 819.72, Bacillus laterosporus, and Bacillus amyloliquefaciens 36,45,52,58 (Figure 4). CCD: Figure 5 shows CCD.…”

“… 51 Also, Bacillus amyloliquefaciens has been shown to use yeast extract (2 g/L), as an organic nitrogen source to produce alpha-amylase. 52 A study on Penicillium notatum IBGE 03 used corn steep as the organic nitrogen source to optimize alpha-amylase production. 42 Soybean has been used as a source of nitrogen by Aspergillus oryzae CBS 819.72, which produced alpha-amylase on optimized condition.…”

A statistical approach for the production of thermostable and alklophilic alpha-amylase from Bacillus amyloliquefaciens KCP2 under solid-state fermentation

“…A marked enhancement in production of alpha-amylase by B. amyloliquefaciens in flask fermentation using statistical methods was reported by Zhao et al ( 2011 ). Use of Plackett–Burman design to screen different nutrients affecting production of thermostable β-amylase and pullulanase by Clostridium thermosulfurogenes SV2 has been reported by Reddy et al ( 1999 ).…”