Preparative purification ofTrichoderma reesei native and “core” cellobiohydrolase I by electrophoresis and chromatofocusing

Abstract

The enzymes present in the cellulase complex produced by the fungus Trichoderma reesei have been the subject of considerable attention due to their potential for converting cellulosic materials into glucose for further use in fermentation processes. Cellobiohydrolase I (CBH I) is the major component of crude commercial fungal cellulase preparations and catalyzes the conversion of insoluble cellulose into cellobiose. We have been interested, recently, in the reduction of native and core'' CBH I (the primary structure of CBH I with a catalytic head region) and have needed to develop a method for their preparative purification. We now report that by using electrophoresis and chromatofocussing, preparative quantities of both native and core'' CBH I have been obtained. Since their pI values are different, milligram quantities of core'' CBH I can be generated and purified from the native enzyme by chromatofocussing within 2 hours. 13 refs., 7 figs., 2 tabs.