Production, Purification and Partial Characterization of Moderately Thermostable β-Amylase from Bacillus polymyxa BWB-01

Title:Production, Purification and Partial Characterization of Moderately Thermostable β-Amylase from Bacillus polymyxa BWB-01

Volume: 4 Issue: 2

Author(s): Folasade M. Olajuyigbe and Folasade M. Adeniyi

Affiliation:

Keywords: β-amylase, Bacillus polymyxa BWB-01, characterization, enzyme, production, purification.

Abstract: Despite the higher activity possessed by bacterial β-amylases, plant β-amylases are widely used for industrial processes. It is crucial that targeted improvements must be made on bacterial β-amylases for enhanced production, thermal and operational stability for their commercial viability. In this study, Bacillus polymyxa BWB-01 was cultivated in submerged fermentation and influence of cultivation conditions (pH, temperature, carbon and nitrogen sources) on β-amylase production was investigated. β-amylase was purified in a three-step procedure before characterization. Maximum β-amylase production (75.88 U/ml) was obtained at pH 6.0 and 40 °C with soluble starch and malt extract as carbon and nitrogen sources. β-amylase was purified 5.64-fold with 19 % yield. The purified enzyme had optimum activity at 50 °C and pH 6.0, and exhibited remarkable stability over broad pH range (4.0 - 8.0) after 60 min incubation having residual activity of about 90% at pH 7.0, 6.0 and pH 5.0 with 50% residual activity at pH 4.0 and 8.0, respectively. The purified β-amylase uniquely retained 80% and 73% of its original activDespite the higher activity possessed by bacterial β-amylases, plant β-amylases are widely used for industrial processes. It is crucial that targeted improvements must be made on bacterial β- amylases for enhanced production, thermal and operational stability for their commercial viability. In this study, Bacillus polymyxa BWB-01 was cultivated in submerged fermentation and influence of cultivation conditions (pH, temperature, carbon and nitrogen sources) on β-amylase production was investigated. β-amylase was purified in a three-step procedure before characterization. Maximum β- amylase production (75.88 U/ml) was obtained at pH 6.0 and 40 °C with soluble starch and malt extract as carbon and nitrogen sources. β-amylase was purified 5.64-fold with 19% yield. The purified enzyme had optimum activity at 50 °C and pH 6.0, and exhibited remarkable stability over broad pH range (4.0 - 8.0) after 60 min incubation having residual activity of about 90% at pH 7.0, 6.0 and pH 5.0 with 50% residual activity at pH 4.0 and 8.0, respectively. The purified β-amylase uniquely retained 80% and 73% of its original activity after prolonged incubation (180 min) at pH 6.0 and 5.0, and still had residual activity of 65% after 300 min. The enzyme was moderately thermostable exhibiting 62% residual activity after 60 min at 50 °C and 54% at 60 °C after 30 min. β-amylase activity was enhanced by Mn2+ and Fe2+ but completely inhibited by Cu2+. B. polymyxa BWB-01 is a good producer of β-amylase with improved characteristics for application in food industry.ity after prolonged incubation (180 min) at pH 6.0 and 5.0, and still had residual activity of 65% after 300 min. The enzyme was moderately thermostable exhibiting 62% residual activity after 60 min at 50 °C and 54% at 60 °C after 30 min. β-amylase activity was enhanced by Mn2+ and Fe2+ but completely inhibited by Cu2+. B. polymyxa BWB-01 is a good producer of β-amylase with improved characteristics for application in food industry.

Cite this article as:

Olajuyigbe M. Folasade and Adeniyi M. Folasade, Production, Purification and Partial Characterization of Moderately Thermostable β-Amylase from Bacillus polymyxa BWB-01, Current Biotechnology 2015; 4 (2) . https://dx.doi.org/10.2174/2211550104666150107234112