Isolation of cold-active alkaline protease Bacillus sp. strains, enzyme production, characterization, determination and their biotechnological applications

In this study, the cold alkaline protease enzymes from Bacillus sp. strains isolated from different environments were produced and characterized. For this purpose, different temperatures and pH values of bacteria and enzymes to produce reproductive capabilities on solid plate were investigated.. Besides, different chemicals and NaCl effects on the enzyme activity was determined.Molecular weight of the enzyme from Bacillus sp. NB-16 was estimated as 49.5 kDa on SDS-PAGE. The optimum activity was obtained at 25ºC and the enzyme was retained it?s activity around 91.32% at the end of 30 minutes between 10-80ºC. The optimum activity was obtained at pH 12.0 The enzyme also presented the alkali-stable properties with a residual activity around 94.14 % at 23ºC between pH 7.0-12.0 for 24 hours. The optimum activity of NB-16 protease has shown 92.14% in the 5% NaCl concentration. ß-mercaptoethanol (%159%), urea (122), IAM (177), PHG (123%), stimulated the enzyme activity significantly however SDS (54%) reduced.According to these result, NB-16 enzyme shows cold- active, psychrotolerant, alkaliphile and alkali-stable characters. Due to these properties NB-16 cold-active protease enzyme has capable of using at especially detergent and bioremediation industry processing and food industry.