Production of nattokinase from Bacillus subtilis-optimization and purification by liquid-liquid extraction

Abstract

Nattokinase is an enzyme with thrombolytic activity that finds a wide range of function in cardiovascular diseases prevention and many biomedical applications. In this study, Bacillus subtilis was isolated and identified from soil and screened for the production of nattokinase enzyme. The characterization results of the purified enzyme showed its optimized parameters like carbon source – dextrin at 100 mg/ml, nitrogen source – peptone at 5 mg/ml, pH 7, temperature- 35oC, pH 7, inhibitor - 1 ml of 10% zinc chloride, activator – 1 ml of 10% EDTA and incubation time - 36 h. The enzyme activity found to be gradually declined at temperature above 40oC. Those effects the production are optimized and the crude enzyme obtained was purified by using 2-liquid- liquid extraction technique. The purified enzyme was characterized and the results were found to be promising for future studies and commercial production.