DOI: 10.36347/sajb.2018.v06i08.007

Pages: 598-605

Fermented foods have been explored for their potentials as main sources of fibrinolytic enzymes. In this study, we did screening for fibrinolytic enzymes-producing bacteria isolated from several Indonesian fermented foods, including black tauco, salted tauco, dadih, cassava tape, brem, salted vegetable, fermented fish, terasi, tempe, oncom, fermented fish, tahu, and tempoyak. Fibrinolytic activity was directly determined by fibrin zymogram and fibrin-plating assays. Crude enzyme from selected isolate producing optimal fibrinolytic activity was produced, purified, and further characterized for its biochemical properties (optimum pH, optimal temperature, protease inhibitor, metal ion, and substrate specificity). Crude enzyme was purified using ammonium sulphate fractionation with 65% saturation, dyalisis using nitrocellulose acetate membrane (cut off 10 kDa), concentration with polyethylene glycol, followed by fractionation and purification using fast protein liquid chromatography with DEAE-Sepharose column. Our results demonstrated that a fibrinolytic enzyme (F7 eluate) was successfully isolated, purified, and characterized from the culture filtrate of TH-5 strain isolated from the black tauco. Spesific activity of F7 eluate was 15.421 U/mg with 45.6 purification fold compared to its crude enzyme. The optimum pH and temperature for enzyme activity of F7 eluate were 7 and 50oC, respectively. Enzyme activity of F7 eluate was totally inhibited by addition of phenylmethylsolfonylfluoride (PMSF) and N-α-tosyl-L-lysine chloromethyl ketone (TLCK) inhibitors at final concentration of 1 mM, indicating that enzyme was grouped in serine protease family. Metal ions (K+, Na+, Mg2+, Mn2+, and Cu2+) at total concentration of 5 mM strongly reduced enzyme activity with residual activity less than 30%, indicating that enzyme catalytic of F7 eluate was significantly inhibited by the addition of the ions. Fibrin zymogram profile of F7 eluate resulted in single fibrinolytic band with est