Study On Purification Of Cellulase From Penicillium Sp. And Pretreatment Of Cellulosic Biomass

Cellulosic biomass is the most abundant renewable resource on the earth. Bagasse is a kind of bulk solid waste of sugar industry which has been received more and more attention recently.The strain Penicillium sp.T24-2 was isolated and preserved in this lab which could produce cellulases at a high level by solid-state fermentation. Two kinds ofβ-glucosidase were isolated by anion exchange DEAE-Sepharose Fast Flow column chromatography from crude enzyme solution. The molecular weights are determined as 20 kDa and 35 kDa by SDS-PAGE, respectively. The optimum temperature for catalysis are determined as 60℃and 45℃, respectively. The optimum pH value are 4.5 and 5.0, respectively.The DNS method which was generally applied in the determination of reducing sugar was analysed in this paper. The research showed that the chemical reagent such as acid, ammonium salt and magnesium salt could interfere the determination result seriously. Analysis of CMCase method showed the dilution of enzyme, temperature, pH, reaction time and concentration of CMC-Na could affect the determination result at different level.After the bagasse was treated with alkali, the lignin, most substance dissolved by neutral detergent and a portion of hemicellulose in the bagasse were removed. The treatment temperature, time and the kind of alkali would greatly affect the saccharification. The higher saccharification yield was obtained when the baggase was treated under alkali with a higher concentration and longer time and appropriate concentration of sodium sulfite. However, the bagaase treated with alkali was not properly for producing cellulase by solid-state fermentation.