Standardization Of Methodology For Extracting Ligninolytic Enzymes From Solid State Fermentation

The recovery of ligninolytic enzymes from fermented wheat bran derived upon growth of Stereum ostrea under optimal conditions was carried out with 20 ml of six different solvents - acetate buffer, phosphate buffer, citrate buffer, lactate buffer, succinate buffer and distilled water. Of all the solvents tested, phosphate buffer appeared to be the appropriate buffer for optimum recovery of all the three enzymes. Optimization of different volumes of phosphate buffer reveals that 25m1 of solvent causes the maximum recovery of all the enzymes. Duration of soaking of fermented bran for recovery of enzymes was optimized by incubating fermented bran with solvent for different intervals - 30, 60 and 120 minutes. Thirty minute duration was optimum for maximum recovery of laccase, MnP and LiP while high protein content was recovered after 2 hours duration.Of the two leaching conditions - shaking and stationary considered in the study, shaking was more effective for recovery of enzymes and proteins. Experiments were further conducted to find out no. of washes required for complete recovery of enzymes from fermented bran. Results of these experiment indicated that the first two washes were sufficient to totally extract the enzyme and protein from the bran.