A novel thiol-dependent serine protease from Neocosmospora sp. N1.

ABSTRACT: Alkaline proteases have several industrial applications. In the present study, newly isolated Neocosmospora sp. N1 was screened as hyper producer of serine protease. A multimeric protease of the fungus was purified to homogeneity till 96.78 fold purification with 22.51% recovery. The homogeneity of purified enzyme was checked by native PAGE and its molecular weight was found to be 198.03 kDa by MALDI-TOF. On SDS-PAGE analysis, enzyme was found to be a hetero oligomer of 17.66 kDa and 20.89 kDa subunits. The purified enzyme showed maximum activity with casein as substrate at 60 °C and pH 8.5. The Km and Vmax values were found to be 0.015 mg/ml and 454.45 U/ml, respectively. The enzyme was completely inhibited by PMSF, while the activity was 40% enhanced using ?-mercaptoethanol, suggesting that it is a thiol-dependent serine protease. The purified protease was active over an alkaline pH range from 7 to 12 and temperatures from 20 °C to 60 °C. The enzyme exhibited excellent stability, almost 100% towards organic solvents such as toluene, benzene and hexane, surfactants such as Triton X-100, Tween-20, Tween-80 and SDS, as well as commercial detergents. The significant properties of purified enzyme assure that it could be a potential candidate for commercial purposes.

SUBMITTER: Matkawala F 

PROVIDER: S-EPMC6699422 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature