Regulation of Adsorption Behavior of Carbohydrate-Binding Module Family 1 and Endo-.BETA.-1,4-Glucanase onto Crystalline Cellulose

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  • Nozaki Kouichi
    Department of Chemistry and Material Engineering, Faculty of Engineering, Shinshu University
  • Nishijima Hiroto
    Department of Chemistry and Material Engineering, Faculty of Engineering, Shinshu University
  • Arai Tsutomu
    Department of Chemistry and Material Engineering, Faculty of Engineering, Shinshu University
  • Mizuno Masahiro
    Department of Chemistry and Material Engineering, Faculty of Engineering, Shinshu University
  • Sato Nobuaki
    Department of Chemistry and Material Engineering, Faculty of Engineering, Shinshu University
  • Amano Yoshihiko
    Department of Chemistry and Material Engineering, Faculty of Engineering, Shinshu University

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  • Regulation of Adsorption Behavior of Carbohydrate-Binding Module Family 1 and Endo-β-1,4-Glucanase onto Crystalline Cellulose

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Family 1 carbohydrate-binding module (CBM1) is widely distributed in fungal cellulases and proteins related to cellulose degradation. It plays an important role in adsorbing proteins and enhancing enzymatic degradation at liquid-solid interfaces. In this study, we constructed a fusion protein (CBM1-GFP) consisting of the CBM1 and a green fluorescent protein, and used this to develop a simple method of regulating CBM1 adsorption onto cellulose, which is available for enzyme recycling. We were able to control the amount of adsorbed CBM1-GFP within the range of 5-65% by increasing the concentration of acetate buffer. The same adsorption behavior was also seen in the recombinant Trametes hirsuta endo-glucanase (rEG) containing the CBM1, but not observed in cellobiohydrolase I (rEx-1). The amount of rEG adsorbed was significantly correlated to its hydrolytic activity against crystalline cellulose. After adsorption in 100 mM acetate buffer, the proteins were desorbed with water at 45°C. At this stage, the amount of adsorbed CBM1-GFP, rEx-1 and rEG decreased to 35, 20 and 7%, respectively. This regulation system can be easily combined with existing saccharification and enzyme recycling processes.

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