| Cellulose is the most abundant renewable organic compound in biomass. Due to energy shortage and air pollution coursed by incomplete combustion of coal, oil and other fossil fuels, the research of cellulose enzymes and the conversion of biomass have become more and more important. The fungi of Trichoderma are the best resource for the extracellular cellulase degradation of cellulose.In this thesis, Trichoderma koningii GIMP3.444 was used for cellulose production. The extracellular cellulose was purified by methods of ammonium sulfate precipitation, DEAE-Sepharose FF anion exchange chromatography and Sephadex G-75 gel filtration chromatography.A chitinase was obtained from Trichoderma koningii which was identified as a single band by SDS-PAGE and its molecular weight was 44.6 kDa. The enzyme both has chitinase and CMCase activity. It can degrade chitin colloid and CMC-Na, the former producing N-acetylglucosamine, and the latter sugar. The kinetic analysis of the chitinase towards CMC-Na showed that its specific activity was 19.65 IU-mg"1. The optimum condition was pH 4.5 and temperature 55℃. Km was 7.22 mg/mL. The enzymatic activity was modified by metal ions, which was significantly inhibited about 35% by Ca2+. The kinetic analysis of the chitinase towards chitin colloid showed that its specific activity was 33.42 IU-mg-1. The optimum condition was pH 4.0 and temperature 50℃. Km was 0.60 mg/mL, while Cu2+, Mn2+, Zn2+can significantly inhibited on the enzyme activity.An endo-β-1,6-galactanase, which can hydrolysis of filter paper and produce sugars, was obtained. Its molecular weight was 51 kDa. Another obtained protein was glucosidase, whose molecular weight was 35.6 kDa.There was a protein which confirmed to be homogeneous on SDS-PAGE with molecular weight of 53 kDa. MALDI-TOFMS analysis indicated that it may be a new protein. This protein can improve cellulase synergistic as much as 112%, but the protein itself does not have any cellulase activity. |
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