Isolation Screening Characterization and Production of Fibrinolytic Enzyme from Streptomyces Althioticus BN22

Abstract

Fibrinolytic enzyme has gained an important place in the treatment of cardiovascular diseases in recent times. In this study, 114 different marine actinomycete strains from Marina beach, Besant nagar beach and Neelankarai beach were isolated from the marine samples. We used the starch casein agar plate method (SCA) to isolate the above mentioned strains. newlineAmongst the 114 actinomycete strains, 40 isolated strains showed protease activities in skim milk agar plate method, and the rest of the isolates did not show the protease activity. The protease producing 40 strains were tested for fibrinolytic enzyme activity by the fibrin plate technique which showed a clear zone of fibrinolytic activity. newlineOut of 40 protease producing actinomycete strains, the fibrinolytic activity was shown in 12 actinomycete isolates only. The highest fibrinolytic enzyme activity strain BN22, was identified for further investigation. The amylase and cellulase activity was studied in 114 strains. Amylase activity by starch agar plate method was detected in 13 strains and cellulase activity by the cellulase plate method was detected in 12 strains. Human pathogenic bacteria like E. coli, P. aeruginosa and S. aureus were tested with the agar well diffusion technique for antimicrobial activity. newlineTwenty-two isolates showed maximum antimicrobial activity against human pathogen bacteria. The potential fibrinolytic strain of Streptomyces althoticus BN22 was identified by 16SrRNA sequencing. Among the six different carbon source tested, glucose at 1% was found suitable for fibrinolytic enzyme production (51,570 Eu/ml). newlineThe optimum pH for maximum production of fibrinolytic enzyme activity was identified as pH7.5 and temperature for the same was identified as 50and#9702;C respectively. Among the seven different nitrogen source tested, yeast extract at 1% favored maximum production of fibrinolytic enzyme (54,625 Eu/ml) followed by soybean and peptone. newlineA double bands in SDS-PAGE was identified and the molecular mass was determined as 58kDa. Further sequences were an