Pilot-scale production and purification of the cellulolytic enzyme system from the white-rot fungus Phanerochaete chrysosporium  [1991]

The fungus Phanerochaete chrysosporium (Sporotrichum pulverulentum) was grown on a pilot scale in a 100-liter automatic/fermenter and enzyme secretion was followed by continuous HPLC monitoring on an analytical Mono Q HR 5/5 column. The fermentation process yielded a culture filtrate with cellulose-degrading enzymes in a concentration several times higher than those reported from shaken-flask cultures. A quick and efficient downstream process involving ammonium sulfate precipitation afforded high recovery of the enzymes. A new purification approach for the cellulolytic enzyme system is also presented. Three 1,4-beta-D-glucan endoglucanases (EC 3.2.1.4) and three 1,4-beta-D-glucan cellobiohydrolases (EC 3.2.1.91) are brought to physicochemical homogeneity from the crude culture filtrate in a simple two- or three-step purification procedure. Ion-exchange, hydrophobic interaction, and gel chromatography are used as the main purification methods. Two of the purified enzymes have not been reported previously in the literature