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高等学校化学研究 ›› 2011, Vol. 27 ›› Issue (4): 557-561.

• Articles • 上一篇    下一篇

Purification and Characterization of β-Glucosidase from a Newly Isolated Strain Tolypocladium cylindrosporum Syzx4

ZHANG Yi-bo1, YUAN Li-jia1, CHEN Zhao-jie2, FU Lu1, LU Jia-hui1, MENG Qing-fan1, HE Huan1, YU Xiao-xiao1, LIN Feng1* and TENG Li-rong1*   

  1. 1. Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, College of Life Science, Jilin University, Changchun 130012, P. R. China;
    2. Department of Endocrinology, Chian-Japan Union Hospital of Jilin University, Changchun 130033, P. R. China
  • 收稿日期:2011-01-10 修回日期:2011-03-02 出版日期:2011-07-25 发布日期:2011-06-29
  • 通讯作者: LIN Feng;TENG Li-rong E-mail:linfeng@jlu.edu.cn; tenglr@jlu.edu.cn
  • 基金资助:

    Supported by the Important Agriculture Program of the Jilin Province Technology Department, China(No.20096013), the  Jilin University Basic Science Research Fund, China(No.200903259), the Graduate Innovation Fund of Jilin University, China (No.20101043) and the Project of Jilin Fuel Alcohol Company Ltd., China.

Purification and Characterization of β-Glucosidase from a Newly Isolated Strain Tolypocladium cylindrosporum Syzx4

ZHANG Yi-bo1, YUAN Li-jia1, CHEN Zhao-jie2, FU Lu1, LU Jia-hui1, MENG Qing-fan1, HE Huan1, YU Xiao-xiao1, LIN Feng1* and TENG Li-rong1*   

  1. 1. Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, College of Life Science, Jilin University, Changchun 130012, P. R. China;
    2. Department of Endocrinology, Chian-Japan Union Hospital of Jilin University, Changchun 130033, P. R. China
  • Received:2011-01-10 Revised:2011-03-02 Online:2011-07-25 Published:2011-06-29
  • Contact: LIN Feng;TENG Li-rong E-mail:linfeng@jlu.edu.cn; tenglr@jlu.edu.cn
  • Supported by:

    Supported by the Important Agriculture Program of the Jilin Province Technology Department, China(No.20096013), the  Jilin University Basic Science Research Fund, China(No.200903259), the Graduate Innovation Fund of Jilin University, China (No.20101043) and the Project of Jilin Fuel Alcohol Company Ltd., China.

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摘要： A higher β-glucosidase-producing strain was isolated and identified as Tolypocladium cylindrosporum syzx4 based on its morphology and internal transcribed spacer(ITS) rDNA gene sequence. The present study is to ferment, purify and characterize a β-glucosidase from T. cylindrosporum gams. The enzyme was purified to homogeneity by sulfate precipitataion, diethylaminoethyl cellulose anion exchange chromatography and Sephadex G-100 gel filtration with a 9.47-fold increase in specific activity and a recovery of 12.27%. The molecular weight(Mw) of the β-glucosidase was estimated to be 58600 by SDS-PAGE, which is much lower than that of the enzyme from other fungi. The β-glucosidase shows high activity over a wide range of temperature from 35 °C to 70 °C and the optimum pH is approximately 2.4. Then the kinetic parameters Km(0.85 mmol/L) and vmax[85.23 mmol/(L·s)] of the β-glucosidase were studied, respectively with high affinity p-nitrophenyl-β-D-glucopyranoside(p-NPG) as the substrate, inhibition constant Ki(39.5 mmol/L) with the tolerance of glucose. Although β-glucosidases have been purified and characterized from several other sources, T. cylindrosporum β-glucosidase with the unique optimal pH and higher tolerance of glucose distinguished from others makes the β-glucosidase a potential application in simultaneous saccharification and fermentation(SSF).

关键词: β-Glucosidase, Purification, Characterization, Thermoacidophilic, Tolypocladium cylindrosporum syzx4

Abstract: A higher β-glucosidase-producing strain was isolated and identified as Tolypocladium cylindrosporum syzx4 based on its morphology and internal transcribed spacer(ITS) rDNA gene sequence. The present study is to ferment, purify and characterize a β-glucosidase from T. cylindrosporum gams. The enzyme was purified to homogeneity by sulfate precipitataion, diethylaminoethyl cellulose anion exchange chromatography and Sephadex G-100 gel filtration with a 9.47-fold increase in specific activity and a recovery of 12.27%. The molecular weight(Mw) of the β-glucosidase was estimated to be 58600 by SDS-PAGE, which is much lower than that of the enzyme from other fungi. The β-glucosidase shows high activity over a wide range of temperature from 35 °C to 70 °C and the optimum pH is approximately 2.4. Then the kinetic parameters Km(0.85 mmol/L) and vmax[85.23 mmol/(L·s)] of the β-glucosidase were studied, respectively with high affinity p-nitrophenyl-β-D-glucopyranoside(p-NPG) as the substrate, inhibition constant Ki(39.5 mmol/L) with the tolerance of glucose. Although β-glucosidases have been purified and characterized from several other sources, T. cylindrosporum β-glucosidase with the unique optimal pH and higher tolerance of glucose distinguished from others makes the β-glucosidase a potential application in simultaneous saccharification and fermentation(SSF).

Key words: β-Glucosidase, Purification, Characterization, Thermoacidophilic, Tolypocladium cylindrosporum syzx4