Effects of Micronutrients on the Production of Heparinase from Bacillus lentus

Dhanalakshmi.R, Manasa.A, Ramesh Babu.N.G

Abstract: Heparinases are lyases capable of recognizing and cleaving different sequences of heparin (a highly sulfated glycosaminoglycan). These enzymes have potential applications in several domains like treatment of Thromboelastography blood samples and removal of interference caused by heparin, monitoring of the alterations in coagulation function, protection against reperfusion injury following ischemia and reduction of restenosis following angioplasty. Neutralization, detection and determination of plasma heparin and quality control in heparin manufacture also involve use of heparinases. In view of the extensive roles played by these enzymes, the current study aims to identify and isolate bacteria capable of producing this enzyme in the in vitro study followed by enzyme extraction and purification. The work dealt with production of enzyme by adding various micronutrients so as to detect the best micronutrient for maximum yield. Once the production is done, the crude enzyme has been extracted and subjected to various purification protocols like ammonium sulfate precipitation, dialysis, ion exchange chromatography, gel filtration, etc. The enzyme assay, optimization and protein concentration estimations were also performed and Bacillus lentus resulted in the maximum yield in the presence of ferric chloride.

Keywords: Heparinases, heparin, Thromboelastography, micronutrients, purification