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Properties of cellulosomal family 9 cellulases from Clostridium cellulovorans

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Abstract

The cellulosomal family 9 cellulase genes engH, engK, engL, engM, and engY of Clostridium cellulovorans have been cloned and sequenced. We compared the enzyme activity of family 9 cellulosomal cellulases from C. cellulovorans and their derivatives. EngH has the highest activity toward soluble cellulose derivatives such as carboxymethylcellulose (CMC) as well as insoluble cellulose such as acid-swollen cellulose (ASC). EngK has high activity toward insoluble cellulose such as ASC and Avicel. The results of thin-layer chromatography showed that the cleavage products of family 9 cellulases were varied. These results indicated that family 9 endoglucanases possess different modes of attacking substrates and produce varied products. To investigate the functions of the carbohydrate-binding module (CBM) and the catalytic module, truncated derivatives of EngK, EngH, and EngY were constructed and characterized. EngHΔCBM and EngYΔCBM devoid of the CBM lost activity toward all substrates including CMC. EngKΔCBM and EngMΔCBM did not lose activity toward CMC but lost activity toward Avicel. These observations suggest that the CBM is extremely important not only because it mediates the binding of the enzyme to the substrates but also because it participates in the catalytic function of the enzyme or contributes to maintaining the correct tertiary structure of the family 9 catalytic module for expressing enzyme activity.

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Acknowledgements

This research was supported in part by grant DE-DDF03-92ER20069 from the U.S. Department of Energy and a grant from the RITE Institute, Kyoto, Japan.

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Author notes

  1. Akihiko Kosugi

    Present address: Japan International Research Center for Agricultural Sciences (JIRCAS), 305-8686, Tsukuba, Ibaraki, Japan

  2. Roger Koukiekolo

    Present address: SIMS Department, 100 Sussex drive, ON K1A 0R6, Ottawa, Canada

Authors and Affiliations

  1. Section of Molecular and Cellular Biology, University of California, Davis, CA, 95616, USA

    Takamitsu Arai, Helen Chan & Roy H. Doi

  2. Research Institute of Innovative Technology for the Earth (RITE), Kyoto, Japan

    Hideaki Yukawa & Masayuki Inui

Authors
  1. Takamitsu Arai
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  2. Akihiko Kosugi
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  3. Helen Chan
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  4. Roger Koukiekolo
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  5. Hideaki Yukawa
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  6. Masayuki Inui
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  7. Roy H. Doi
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Correspondence to Roy H. Doi.

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Arai, T., Kosugi, A., Chan, H. et al. Properties of cellulosomal family 9 cellulases from Clostridium cellulovorans . Appl Microbiol Biotechnol 71, 654–660 (2006). https://doi.org/10.1007/s00253-005-0249-6

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  • DOI: https://doi.org/10.1007/s00253-005-0249-6

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