Purification and characterization of thermoactive serratiopeptidase from Serratia marcescens AD-W2

AMB Express

Md. Mehedi Hossain¹  Mohd Jamal Dar¹  Jasmine Kour Khosla²  Diksha Koul³  Devtulya Chander³  Asha Chaubey³ 

[1] Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, 180001, Jammu, India;Academy of Scientific and Innovative Research, CSIR-Human Resource Development Centre, Campus Ghaziabad, 201002, Ghaziabad, India;Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, 180001, Jammu, India;Fermentation Technology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, 180001, Jammu, India;Academy of Scientific and Innovative Research, CSIR-Human Resource Development Centre, Campus Ghaziabad, 201002, Ghaziabad, India;

关键词: Serratia marcescens;    Metalloprotease;    Serratiopeptidase;    Serralysin;    Homology modelling;   

DOI  :  10.1186/s13568-021-01215-7

来源: Springer

【 摘 要 】

Serratiopeptidase is a proteolytic enzyme extensively used as an anti-inflammatory and analgesic drug. Present work reports a thermoactive serratiopeptidase from Serratia marcescens AD-W2, a soil isolate from the North-Western Himalayan region of India. The extracellular metalloprotease has been purified by a simple two-step procedure resulting in a specific activity of 20,492 Units/mg protein with 5.28-fold purification. The molecular mass of the metalloprotease, as determined by SDS-PAGE was ~ 51 kDa. The purified serratiopeptidase presented optimum activity at pH 9.0, temperature 50 °C and stability in wide pH and temperature range. Critical temperature of 50 °C confirmed the thermoactivity of the purified serratiopeptidase. The kinetic studies of the purified serratiopeptidase revealed Vmax and Km of 57,256 Units/mL and 1.57 mg/mL, respectively, for casein. The purified serratiopeptidase from S. marcescens AD-W2 was found to be 100% identical to serralysin from Serratia marcescens ATCC 21074/E-15. The catalytic domain comprising of Zn coordinated with three histidine residues (His192, His196, His202), along with glutamate (Glu193) and tyrosine (Tyr232) residues, further confirmed that the purified protein is identical to serralysin.

【 授权许可】

CC BY