Isolation of cold active alkaline amylase producing Bacillus sp. strains, enzyme production, characterization and investigation of industrial applications

In this study, the amylase enzyme from Bacillus sp. strains isolated from Ankara and salt lake was produced and characterized. For this purpose, optimum temperature and pH value, temperature and pH stability, SDS PAGE zymogram analysis and thin layer chramatography analysis of enzyme was investigated.Molecular weight of the enzyme from Bacillus sp. NCA35 was estimated as 205 kDa on SDS-PAGE. The enzyme showed its optimum activity at 25ºC and pH 8.0. The enzyme activity was retained around 96% of the original activity within 30 minutes between 10-40ºC. Enzyme was also stable about %53 at 25oC for 24 hours. The residual enzyme activity was 56% in the presence of 3% NaCl at 60 min.As the ß-mercaptoethanol stimulated the enzyme activity (103%), BaCl2, EDTA, CaCl2, NaCl, ZnCl2, MnCl2 (5mM), PMSF (3mM) and Urea (8M) inhibited the activity to %27, %42, %35, %43, %40 %36, %41 and %58, respectively. Glucose, maltose and maltose-derived oligosaccharide were determined as hydrolysis products by thin layer chromatography.According to these results, NCA35 enzyme shows cold active, acido-alkaline, thermostable and pH-stable properties. Therefore, NCA35 cold active amylase enzyme has the potentials to use in detergents, beverage and food industries, bioremediation applications and prebiotic production.