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《Journal of Harbin Institute of Technology》 2006-11
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Purification and characteristics of celluloses from trichoderma aureoviride

SONG Jin-zhu,YANG Qian,CHEN Zhong-xiang,ZHANG Jing(Dept.of life science and bioengineering,Harbin Institute of Technology,Harbin 150001,China,)  
Three endoglucanases and two β-glucosidases from the fungus were separated and purified to electrophoretic homogeneous by(NH_4)_2SO_4 gradient precipitation,Sephadex G-100 gel chromatography and DEAE Sephadex A-50 ion exchange chromatography.They were designated to be EGⅠ,EGⅡ,EGⅢ,BGⅠ and BGⅡ.Five components had molecular weights of 62.3 ku,71.9 ku,52.6 ku,85.3 ku and 78.3 ku by SDS-PAGE,respectively,and IEF showed their isoelectric point were 5.4,4.8,5.0,5.6,and 5.8,respectively.All of them had maxium reactive activities at pH 5.The optimum temperature of endoglucanases is 60 ℃,and the optimum temperature of β-glucosidases is 55 ℃.The activities of all components were stimulated by Mn~(2+),Fe~(2+),Zn~(2+),and especially,EG were mostly activated by Mn~(2+),while the activities of all components were inhibited by Mg~(2+),Cu~(2+),carbamide.The kinetics showed the K_m of five components were 0.093 mg/ml,0.083 8 mg/ml,0.079 0 mg/ml,0.085 1 mg/ml and 0.077 8 mg/ml,respectively. K_(cat) were 101.7 s~(-1)、119.6 s~(-1),11.4 s~(-1),111.4 s~(-1) and 162.1 s~(-1),respectively.There was no relativies between the catalytic efficiency of endoglucanases and its K_m,however,there was correlation between the catalytic efficiency of β-glucosidases and its K_m. Substrate specificity of the five components measurement indicated that BGⅠ and BGⅡ could only active Salicin,yet EGⅠ,EGⅡ and EGⅢ could hydrolyse not only CMC-Na,but also filter paper and straw powder faintly.
【Fund】: 国家自然科学基金资助项目(50178021)
【CateGory Index】: Q814.1
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