Purification and characterization of novel thermostable and Ca-independent α-amylase produced by Bacillus amyloliquefaciens BH072.

In the present study, a novel α-amylase produced by Bacillus amyloliquefaciens BH072 was purified and characterized. The molecular weight of purified α-amylase was approximately 68 kDa, determined by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and ten amino acid of N-terminal was NSGLNGYLTH. The kinetic parameters Km and Vmax were 4.27 ± 0.21 mg/mL and 987.34 ± 23.34 U/mg, respectively. Purified α-amylase showed maximal activity at pH 7 and 60 °C. Enzyme remained stable in pH range 6.0-7.0 and 50-80 °C. The activity of the α-amylase was Ca2+ independent and stability in the presence of surfactant, oxidizing and bleaching agents. The β-mercaptoethanol and EDTA greatly enhanced and reduced α-amylase activity, respectively. This enzyme has high hydrolysis rate toward corn, wheat and potato starch and hydrolyzes soluble starch to glucose, maltose, maltotriose and maltotetraose, indicating that the α-amylase represents a promising candidate for applications in the food industry.