utilization of red lentil flour substrate for the production of nattokinase enzyme by bacillus subtilis natto

In this research, the optimal production conditions of nattokinase using bacillus subtilis natto on red lentil flour substrate were studied. to optimize the fermentation process, the effects of three variables of fermentation time (24, 48 and 72 h), molasses concentration (3, 5 and 7%) and water content (50, 75 and 100 ml) on fibrinolytic and protease activities were investigated. the central composite design (ccd) was employed, and the results were modelled and analyzed using response surface methodology (rsm). coefficients of determination, r2, of fitted regression models for fibrinolytic (quadratic model) and protease (linear model) activities were 97.15 and 90.12%, respectively, and their lack-of-fit was not significant at 95%. hence, the models for all the responses were highly adequate. fibrinolytic activity increased significantly (p≤0.05) with increasing water content and fermentation time, while increasing the amount of molasses decreased fibrinolytic activity (p≤0.05). with increasing the amount of water, protease activity also increased, but this increase was not significant, while increasing the amount of molasses and fermentation time led to a significant decrease and increase in protease activity (p≤0.05), respectively. concerning optimization, the optimal fermentation conditions were determined as 92.38 ml of water, 3.66% molasses, and 70.90 h of fermentation. under these conditions, the activities of fibrinolytic enzymes and proteases were predicted to be 2476.03 and 1.68 u/g, respectively. for validation of the model, the optimal sample was produced, and the experimental responses were compared with the responses predicted by the model. the experimental values obtained were quite close to those predicted by the model, indicating the validity of the model. the results of this study showed that red lentil could be used as a substrate for fermentation by b. subtilis to produce the enzyme nattokinase. 

utilization of red lentil flour substrate for the production of nattokinase enzyme by bacillus subtilis natto

in this research, the optimal production conditions of nattokinase using bacillus subtilis natto on red lentil flour substrate were studied. to optimize the fermentation process, the effects of three variables of fermentation time (24, 48 and 72 h), molasses concentration (3, 5 and 7%) and water content (50, 75 and 100 ml) on fibrinolytic and protease activities were investigated. the central composite design (ccd) was employed, and the results were modelled and analyzed using response surface methodology (rsm). coefficients of determination, r2, of fitted regression models for fibrinolytic (quadratic model) and protease (linear model) activities were 97.15 and 90.12%, respectively, and their lack-of-fit was not significant at 95%. hence, the models for all the responses were highly adequate. fibrinolytic activity increased significantly (p≤0.05) with increasing water content and fermentation time, while increasing the amount of molasses decreased fibrinolytic activity (p≤0.05). with increasing the amount of water, protease activity also increased, but this increase was not significant, while increasing the amount of molasses and fermentation time led to a significant decrease and increase in protease activity (p≤0.05), respectively. concerning optimization, the optimal fermentation conditions were determined as 92.38 ml of water, 3.66% molasses, and 70.90 h of fermentation. under these conditions, the activities of fibrinolytic enzymes and proteases were predicted to be 2476.03 and 1.68 u/g, respectively. for validation of the model, the optimal sample was produced, and the experimental responses were compared with the responses predicted by the model. the experimental values obtained were quite close to those predicted by the model, indicating the validity of the model. the results of this study showed that red lentil could be used as a substrate for fermentation by b. subtilis to produce the enzyme nattokinase.