Streptokinase Enzyme Lab Report

ISOLATION, IDENTIFICATION OF STREPTOKINASE PRODUCING STREPTOCOCCUS SPECIES AND PRODUCTION OF STREPTOKINASE ABSTRACT:

The objective of the study is to identify a potent streptococcal species producing streptokinase enzyme from different samples. Various food samples and soil samples were collected and analyzed for the potent streptococcal strain. They were confirmed for streptococcus species by biochemical characterization. Further, they were screened for the streptokinase activity. Among them curd sample and bore soil sample showed good activity and they were taken for further analysis and production process. They were compared based on their activity of clot lysis and casein digestion. The isolates of curd sample were -hemolytic streptococci

The selected colonies were streaked onto blood agar plates for observing the hemolytic patterns. The potent strains were selected for biochemical characterization. These biochemical tests were performed in reference to Cowan and Steel’s Manual for bacterial identification (Holt et al., 1993).

(a) (b)

Fig 1 Gram staining showing gram positive cocci in chains; (a) bore soil (b) curd

PRODUCTION OF STREPTOKINASE (SK):

The potent strains producing streptokinase were cultured in 100mL of the production medium (g/100 mL: Glucose - 0.5 g, Yeast Extract - 0.5 g, KH2PO4 - 0.25 g, MgSO4·7H2O 0.04 g, NaHCO3 - 0.1 g, CH3COONa·3H2O - 0.1 g, FeSO4·7H2O - 0.002 g, MnCl2·4H2O - 0.002 g, pH 7.5) and were kept for incubated at 37C for 24 h (Baewald et al., 1975). Once incubation is done, the individual cultures were centrifuged at 10,000 rpm for 30 mins. The cell-free supernatant was passed through 0.45 µm cellulose acetate filter and the filtrates were considered a crude enzyme.

ENZYME ACTIVITY – CASEIN DIGESTION METHOD:

The potent strains were inoculated in Todd- Hewitt broth and kept for incubation at 37ºC for 18-24 hours. The culture was centrifuged (a)curd (b)bore soil

Fig 3 Casein digestion – Enzyme