Purification and Biochemical Characterization of Novel Fibrinolytic Protease Produced by Mesophilic Bacillus Circulans Gd25

A novel fibrinolytic protease from Bacillus circulans GD25 was isolated and characterized for the fibrinolytic activity. The fibrinolytic protease was purified by ammonium sulphate fractionation (70%) and subjected to chromatographic methods like Sephadex G-50, DEAE –Sephadex A-50 columns. The purified enzyme has an approximately 38 kDa in size by SDS-PAGE and gel filtration. Optimum activity was at 35oC and the enzyme was highly active over a wide range of pH from 7.0-9.0 with an optimum at pH 8.0. It exhibited stronger fibrinolytic protease activity. The activity was slightly enhanced by Ca and Mg whereas Zn Cu Co and Fe suppressed the activity. The fibrinolytic protease activity was totally lost in the presence of PMSF, suggesting that the purified enzyme is a serine metallo-protease with potential application in thrombolytic therapy INTRODUCTION: Proteases are proteolytic enzymes which catalyze the hydrolysis of proteins based upon their structures or properties of the active site. There are several kinds of proteases such as serine, metallo, carboxyl, acidic, neutral and alkaline proteases. Proteases are industrially important due to their wide applications in leather processing, detergent industry, food industries, pharmaceutical and textile industry etc 1, 2 . Proteases are obtained from plants, animal organs and microorganisms, with the majority obtained from microbial sources. Currently, a large proportion of commercially available proteases are derived from bacteria and fungi. QUICK RESPONSE CODE DOI: 10.13040/IJPSR.0975-8232.4(12).4559-66 Article can be accessed online on: www.ijpsr.com DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.4(12).4559-66 Selection of the right organism plays a key role in obtaining high yield of desirable enzymes. On the other hand, it is a well-known fact that extra cellular proteases production in a microorganism is greatly influenced by media components, especially carbon and nitrogen sources and metal ions along with other growth parameters. Cardio vascular such as high blood pressure, acute myocardial infarction, ischemic heart disease, valvular heart disease, peripheral vascular disease, arrhythmias, stroke etc. are the primary causes of death. CVDs are the number one cause of death globally: more people die annually from CVDs than from any other cause 3 . Throughout the world, according to data of World Health Organization (WHO) an estimated 17.3 million people died from CVDs in 2008, representing 30% of all global deaths 3 . Of these deaths, an estimated 7.3 million were due to coronary heart disease and 6.2 million were due to stroke 4 .