PURIFICATION AND CHARACTERIZATION OF GLYCEROL PHOSPHATE DEHYDROGENASE PRODUCED FROM WHEAT BRAN USING A MIXED CULTURE SYSTEM

Glycerol phosphate dehydrogenase was produced from wheat bran by the mixed culture fermentation technique. The process involved saccharification of cellulose to glucose by Trichoderma viride followed by conversion of glucose into glycerol by Saccharomyces cerevisiae Y-1347. Different purification steps were applied to the culture filtrate to obtain a pure enzyme preparation. The pure preparation indicated that the molecule consists of one peptide chain, with a molecular weight of 51 000 and isoelectric point of 5.7. The amino acid content was also studied. Lineweaver-Burk analysis gave a k(m) value of 0.033 mmol and V-max of 83.3 mmol/ml/min. The enzyme showed its maximum activity at pH 6.0 when incubated at 25 degrees C for 10 min.