Partial purification and characterization of thermostable,alkaline and chelatorresistant protease from a newly isolated bacillus sp. CY7 and its potential applications in various industries

A thermophile alkaline protease producing bacillus sp. strains isolated from soil samples. enzyme synthesis occurred at temperatures between 20°c and 60°c with an optimum of 55°c and between the ph range of 6.0-13.0 with an optimum ph of 10.0 on skimmilk agar plates. analyses of molecular mass of the partially purified enzyme was carried out by sds-page which revealed two bands as 112 and 97 kda. the enzyme was active in a broad temperature range between 30°c and 100°c,with an optimum at 70°c,and maximum activity was at ph 11.0. the enzyme also presented the alkaline-stable properties with a remaining activity around 94%,at ph 6.0-13.0 for 24 h,at 55°c. the enzyme activity was highly stable between 30-100°c with a remaining activity 91%,after pre-incubation for 1 h. enzyme was exposured to various nacl concentrations (3-30%),and the highest residual activity was determined in the presence of 3% nacl (87%). while after being exposed with β-mercaptoethanol (1%),pmsf (3mm),tween20 (0.1%),tween80 (0.1%),sds (1%),tritonx100 (1%),h2o2 (1%),mncl2 (5mm),cacl2 (5mm),bacl2 (5mm),mgcl (5mm),fecl2 (5mm) and edta (5mm),the enzyme exhibited the following activities 105%,101%,107%,119%,72%,99%,98%,98%,99%,98%,97%,97%,96% and 99%,respectively. according to these results,cy7 protease shows thermostable,high alkaline,alkali-stable and chelator resistant properties. owing to its mentioned properties this protease is an ideal choice for application in detergent formulations,leather and textile industries.