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Heterologous Expression and Characterization of an Endoglucanase from a Symbiotic Protist of the Lower Termite, Reticulitermes speratus

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Abstract

RsSymEG, an endoglucanase of glycosyl hydrolase family (GHF) 7 encoded by a transcript isolated from the symbiotic protist of the termite Reticulitermes speratus, is expressed in Aspergillus oryzae. Interestingly, purified RsSymEG1 has a relatively higher specific activity (603 μmol min−1 mg−1 protein) and V max value (769.6 unit/mg protein) than previously reported data for GHF7 endoglucanase of Trichoderma ressei. It also has the same K m value (1.97 mg/ml) with Clostridium cellulolyticum enzymes that contain cellulose binding module, a property indicative of high affinity to substrate, though no cellulose binding module is found within it. Thin-layer chromatography analysis revealed that RsSymEG1 preferentially hydrolyzes the β-1,4-cellulosic linkage of cellodextrins into cellobiose and glucose.

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References

  1. Sanderson, K. (2006). Nature, 444, 673–676. doi:10.1038/444673a.

    Article  CAS  Google Scholar 

  2. Yamada, A., Inoue, T., Wiwatwitaya, D., et al. (2005). Ecological Research, 20, 453–460. doi:10.1007/s11284-005-0062-9.

    Article  Google Scholar 

  3. Nakashima, K., Watanabe, H., & Azuma, J.-I. (2002). Cellular and Molecular Life Sciences, 59, 1554–1560. doi:10.1007/s00018-002-8528-1.

    Article  CAS  Google Scholar 

  4. Nakashima, K., Watanabe, H., Saitoh, H., et al. (2002). Insect Biochemistry and Molecular Biology, 32, 777–784. doi:10.1016/S0965-1748(01)00160-6.

    Article  CAS  Google Scholar 

  5. Breznak, J. A., & Brune, A. (1994). Annual Review of Entomology, 39, 453–487. doi:10.1146/annurev.en.39.010194.002321.

    Article  CAS  Google Scholar 

  6. Ohtoko, K., Ohkuma, M., Moriya, S., et al. (2000). Extremophiles, 4, 343–349. doi:10.1007/s007920070003.

    Article  CAS  Google Scholar 

  7. Watanabe, H., Nakashima, K., Saito, H., et al. (2002). Cellular and Molecular Life Sciences, 59, 1983–1992. doi:10.1007/PL00012520.

    Article  CAS  Google Scholar 

  8. Li, L., Fröhlich, J., Pfeiffer, P., et al. (2003). Eukaryotic Cell, 2, 1091–1098. doi:10.1128/EC.2.5.1091-1098.2003.

    Article  CAS  Google Scholar 

  9. Inoue, T., Moriya, S., Ohkuma, M., et al. (2005). Gene, 349, 67–75. doi:10.1016/j.gene.2004.11.048.

    Article  CAS  Google Scholar 

  10. Todaka, N., Moriya, S., Saita, K., et al. (2007). FEMS Microbiology Ecology, 59, 592–599. doi:10.1111/j.1574-6941.2006.00237.x.

    Article  CAS  Google Scholar 

  11. Zhou, X., Smith, J. A., Oi, F. M., et al. (2007). Gene, 395, 29–39. doi:10.1016/j.gene.2007.01.004.

    Article  CAS  Google Scholar 

  12. Archer, D. B., & Peberdy, J. F. (1997). Critical Reviews in Biotechnology, 17, 273–306. doi:10.3109/07388559709146616.

    Article  CAS  Google Scholar 

  13. Kitamoto, K. (2002). Advances in Applied Microbiology, 51, 129–153. doi:10.1016/S0065-2164(02)51004-2.

    Article  CAS  Google Scholar 

  14. Mabashi, Y., Kikuma, T., Maruyama, J., et al. (2006). Bioscience, Biotechnology, and Biochemistry, 70, 1882–1889. doi:10.1271/bbb.60052.

    Article  CAS  Google Scholar 

  15. Sasaguri, S., Maruyama, J., Moriya, S., et al. (2008). Applied Microbiology, 54, 343–351.

    Article  CAS  Google Scholar 

  16. Inoue, T., Murashima, K., Azuma, J.-I., et al. (1997). Journal of Insect Physiology, 43, 235–242. doi:10.1016/S0022-1910(96)00097-2.

    Article  CAS  Google Scholar 

  17. McIlvaine, T. C. (1921). The Journal of Biological Chemistry, 49, 183–186.

    CAS  Google Scholar 

  18. Ståhlberg, J., Divne, C., Koivula, A., et al. (1996). Journal of Molecular Biology, 264, 337–349. doi:10.1006/jmbi.1996.0644.

    Article  Google Scholar 

  19. Henrissat, B. (1991). The Biochemical Journal, 280, 309–316.

    CAS  Google Scholar 

  20. Henrissat, B., & Bairoch, A. (1996). The Biochemical Journal, 316, 695–696.

    Google Scholar 

  21. Divne, C., Ståhlberg, J., Teeri, T. T., et al. (1998). Journal of Molecular Biology, 275, 309–325. doi:10.1006/jmbi.1997.1437.

    Article  CAS  Google Scholar 

  22. Kleywegt, G. J., Zou, J. Y., Divne, C., et al. (1997). Journal of Molecular Biology, 272, 383–397. doi:10.1006/jmbi.1997.1243.

    Article  CAS  Google Scholar 

  23. Ward, M., Lin, C., Victoria, D. C., et al. (2004). Applied and Environmental Microbiology, 70, 2567–2576. doi:10.1128/AEM.70.5.2567-2576.2004.

    Article  CAS  Google Scholar 

  24. Van Ardell, J. N., Kwok, S., Schweickart, V. L., et al. (1987). Biotechnology, 5, 60–64. doi:10.1038/nbt0187-60.

    Article  Google Scholar 

  25. Kitamoto, N., Go, M., Shibayama, T., et al. (1996). Applied Microbiology and Biotechnology, 46, 538–544. doi:10.1007/s002530050857.

    Article  CAS  Google Scholar 

  26. Kwon, I., Ekino, K., Got'o, M., et al. (1999). Bioscience, Biotechnology, and Biochemistry, 63, 1714–1720. doi:10.1271/bbb.63.1714.

    Article  CAS  Google Scholar 

  27. Okada, G., & Nishizawa, K. (1975). Journal of Biochemistry, 78, 297–306.

    CAS  Google Scholar 

  28. Okada, G. (1976). Journal of Biochemistry, 80, 913–922.

    CAS  Google Scholar 

  29. Fierobe, H. P., Gaudin, C., Belaich, A., et al. (1991). Journal of Bacteriology, 173, 7956–7962.

    CAS  Google Scholar 

  30. Fierobe, H. P., Bagnara-Tardif, C., Gaudin, C., et al. (1993). European Journal of Biochemistry, 217, 557–565. doi:10.1111/j.1432-1033.1993.tb18277.x.

    Article  CAS  Google Scholar 

  31. Scrivener, A. M., & Slaytor, M. (1994). Insect Biochemistry and Molecular Biology, 24, 223–231. doi:10.1016/0965-1748(94)90001-9.

    Article  CAS  Google Scholar 

  32. Watanabe, H., Nakamura, M., Tokuda, G., et al. (1997). Insect Biochemistry and Molecular Biology, I, 305–313. doi:10.1016/S0965-1748(97)00003-9.

    Article  Google Scholar 

  33. Lynd, L. R., Weimer, P. J., van Zyl, W. H., et al. (2002). Microbiology and Molecular Biology Reviews, 66, 506–577. doi:10.1128/MMBR.66.3.506-577.2002.

    Article  CAS  Google Scholar 

  34. Qi, Meng, Jun, H., & Forsberg, C. W. (2008). The Journal of Bacteriology, 190, 1976–1984. doi:10.1128/JB.01667-07.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was partly supported by the New Energy and Industrial Technology Development Organization (NEDO) program for the development of elemental technology for bioenergy conversion. Partial support was also provided by the Eco-molecular Research Program (RIKEN), the Bio-architect Research Program (RIKEN), and the Program for the Promotion of Basic Research Activity for Innovative Biosciences (PROBRAIN). The work described here was conducted as part of a series of studies under the 2005–2006 UNESCO Postgraduate Inter-university Training Course in Biotechnology sponsored by the Japanese Ministry of Education, Culture, Sports, Science and Technology and the Japanese National Commission for UNESCO.

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Author notes

  1. Crisanto M. Lopez

    Present address: Department of Infection Biology, Leibniz Institute for Natural Product Research and Infection Biology (Hans-Knoell Institute), Beutenbergstrasse 11a, 007745, Jena, Germany

  2. Toshiaki Kudo

    Present address: Faculty of Fisheries, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki, 852-8521, Japan

Authors and Affiliations

  1. Laboratory of Environmental Molecular Biology, RIKEN, 1-7-29 Suehiro-cho, Tsurumi-ward, Kanagawa, 230-0045, Japan

    Nemuri Todaka, Tetsushi Inoue, Kanako Saita, Toshiaki Kudo & Shigeharu Moriya

  2. Laboratory of Environmental Molecular Biology, Graduate School of Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ward, Kanagawa, 230-0045, Japan

    Nemuri Todaka, Tetsushi Inoue, Kanako Saita, Toshiaki Kudo & Shigeharu Moriya

  3. Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan

    Crisanto M. Lopez, Jun-ichi Maruyama, Manabu Arioka & Katsuhiko Kitamoto

  4. TOYOTA Central R&D Labs Inc, 41-1, Yokomichi, Nagakute, Nagakute-cho, Aichi, 480-1192, Japan

    Nemuri Todaka

  5. Biosphere Oriented Biology Research Unit, RIKEN Advanced Science Institute, 1-7-29 Suehiro-cho, Tsurumi-ward, Kanagawa, 230-0045, Japan

    Shigeharu Moriya

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  1. Nemuri Todaka
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Correspondence to Shigeharu Moriya.

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Todaka, N., Lopez, C.M., Inoue, T. et al. Heterologous Expression and Characterization of an Endoglucanase from a Symbiotic Protist of the Lower Termite, Reticulitermes speratus . Appl Biochem Biotechnol 160, 1168–1178 (2010). https://doi.org/10.1007/s12010-009-8626-8

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  • DOI: https://doi.org/10.1007/s12010-009-8626-8

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