Exploration of effective fibrinolytic enzyme from newly isolated alcaligenes aquatilis pjs 1

Abstract

This research focuses on the isolation and identification of an organism from slaughter house soil samples for enzymatic fibrinolytic agent production. this research work differs from previous works in this area as i have used a newly isolated organism alcaligenes aquatilis pjs_1. newlinethe various soil samples were taken from slaughter houses located in various parts of karur district, tamil nadu, india. fibrinolytic enzyme synthesising bacterium was screened using serial dilution technique and chracterized by morphological analysis and biochemical tests (casein hydrolysis, fibrin hydrolysis, indole test, catalase test, methyl red test, voges proskauer test and citrate utilization test). 16s rrna sequencing confirmed the organism as alcaligenes aquatilis pjs_1. the fibrinolytic enzyme production medium was optimized for submerged fermentation by various factors like different energy sources (carbon and nitrogen sources and mineral salts), agitation speed, inoculum age, inoculum volume, temperature and ph. similarly, the culture conditions for solid state fermentation (carbon and nitrogen sources, mineral salts, moisture content and temperature) were optimized with sugarcane bagasse as a production substrate. bioreactor used in the experiment was designed with suitable parameters for effective production of fibrinolytic enzyme by plackett-burman design (pbd) and response surface methodology (rsm). fibrinolytic enzyme produced from bioreactor was purified by gel filtration chromatography. blood clotting assay was performed to determine its anticoagulant property. potential fibrinolytic enzyme producing bacterium was isolated and identified as alcaligenes aquatilis pjs_1 based on the results of biochemical tests and 16s rrna sequencing. optimization of the production medium showed that fructose and urea were an ideal carbon and nitrogen sources at ph 7.0, incubated for 24 hrs at 37ºc. the crude enzyme was purified by acetone precipitation followed by gel filtration chromatography. the enzyme was purified to 1.1 fold and