Bioconversion of agro-industrial wastes for the production of
fibrinolytic enzyme from Bacillus halodurans IND18: Purification and
biochemical characterization
Background: Agro-wastes were used for the production of fibrinolytic
enzyme in solid-state fermentation. The process parameters were
optimized to enhance the production of fibrinolytic enzyme from
Bacillus halodurans IND18 by statistical approach. The fibrinolytic
enzyme was purified, and the properties were studied. Results: A
two-level full factorial design was used to screen the significant
factors. The factors such as moisture, pH, and peptone were
significantly affected enzyme production and these three factors were
selected for further optimization using central composite design. The
optimum medium for fibrinolytic enzyme production was wheat bran medium
containing 1% peptone and 80% moisture with pH 8.32. Under these
optimized conditions, the production of fibrinolytic enzyme was found
to be 6851 U/g. The fibrinolytic enzyme was purified by 3.6-fold with
1275 U/mg specific activity. The molecular mass of fibrinolytic enzyme
was determined by sodium dodecyl sulphate polyacrylamide gel
electrophoresis, and it was observed as 29 kDa. The fibrinolytic enzyme
depicted an optimal pH of 9.0 and was stable at a range of pH from 8.0
to 10.0. The optimal temperature was 60\ub0C and was stable up to
50\ub0C. This enzyme activated plasminogen and also degraded the
fibrin net of blood clot, which suggested its potential as an effective
thrombolytic agent. Conclusions: Wheat bran was found to be an
effective substrate for the production of fibrinolytic enzyme. The
purified fibrinolytic enzyme degraded fibrin clot. The fibrinolytic
enzyme could be useful to make as an effective thrombolytic agent