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1 pp. GH GH GH β- - D- β- β- β- - EC J Alkalitolerant mechanism and improvement of alkalitolerancy of glycoside hydrolases Satoshi Nakamura Department of Bioengineering, Tokyo Institute of Technology, J Nagatsuta-cho, Midoriku, Yokohama, Japan ph ph ph

2 GH http: fam acc_gh.html GH GH GH GH GH GH GH X GH http: thornton-srv databases pdbsum GH GH β α GH β- GH GH GH GH GH GH pka ph pka ph pka GH Bacillus sp. M- M- J ph GH CBM XBD J J Glu Glu J Glu pka X J PDB ID DCK DCJ GH GH pi ph β α Glu Glu ph β- Glu Glu

3 GH GH pka J GH β- β J XBD ph J XBD J J J Asp Glu Glu Arg Lys Lys Asn Gln XBD XBD ph ph ph

4 GH Bacillus sp. M- XBD Bacillus agaradhaerens Xyn Bacillus subtilis Xyn X Dictyoglomus thermophilum XynB Streptmyces sp. S Xyn Aspergillus kawachii XynC Aspergillus niger XYLA I His -Asp His -Asp His -Asp Asp -Lys Glu -Arg Glu -Lys Arg -Glu Glu -Lys Asp -Lys Glu -Arg Glu -Lys Arg -Glu Glu -Lys Asp -Lys Glu -Arg Glu -Lys Arg -Glu Glu -Lys Glu -Arg Asp -Lys Glu -Lys His -Glu Lys -Glu Glu -Lys His -Glu Lys -Glu Glu -Lys His -Glu Lys -Glu Glu -Lys Arg -Asp Glu -His Arg -Asp Arg -Asp Glu -His Arg -Glu Glu -His Arg -Glu J A B Asp Glu Glu Arg Lys Lys Glu Glu J Glu pka Glu pka Lys Lys Arg Lys Arg XBDK R XBD ph ph GH ph ph XBDK R Lys Arg XBDK R Lys Arg

5 XBD ph J XBD ph XBD Arg ph J XBD XBDK R Arg XBDR ph Arg J Arg Lys Trichoderma reesei GH II Ser Thr Arg Ser Thr Arg ph ph

6 J Arg Arg Ser Thr Ser Thr Asn Asn Asn Lys J Arg T. reesei II XBD Ser Thr Arg XBDR Ser Thr Asn Asn Asn Arg ph Arg Lys Arg XBDR K R ph ph XBDR XBDR K R Arg J J GH Thermotoga maritima MSB GH A B A B T. maritima MSB B PCR B B ph ph Glu Glu T. maritima MSB GH B B PCR B B Asn Tyr, Tyr Phe Asn Asp Asn Tyr Asn ph XynTBMutant Asn Asp XynTBN D B XynTB XynTBN D B X PDB ID VBR VBU B GH β α B

7 B XynTB XynTBMutant XynTBN D ph XynTBN D B B GH GH T. maritima MSB B B Asn Tyr Asn Glu Glu Asn Tyr Asn Glu Glu Asn XynTBN D XynTBN D B GH GH ph X Ihsanawati

8 Wong, K.K.Y., Tan, L.U.L., & Saddler, J.N. Microbiol. Rev.,,. Kulkarni, N., Shendye, A., & Rao, M. FEMS Microbiol. Rev.,,. Collins, T., Gerday, C., & Feller, G. FEMS Microbiol. Rev.,,. Nakamura, S. Catal. Surv. Asia,,. Sakata, T., Miyakubo, H., Osada, Y., Wada, R., Takahashi, H., Yatsunami, R., Fukui, T., & Nakamura, S. J. Appl. Glycosci.,,. Umemoto, H., Ihsanawati, Inami, M., Yatsunami, R., Fukui, T., Kumasaka, T., Tanaka, N., & Nakamura, S. Nucleic Acids Symp. Ser.,,. Umemoto, H., Ihsanawati, Inami, M., Yatsunami, R., Fukui, T., Kumasaka, T., Tanaka, N., & Nakamura, S. Biosci. Biotechnol. Biochem.,,. Torronen, A. & Rouvinen, J. Biochemistry,,. Fushinobu, S., Ito, K., Konno, M., Wakagi, T., & Matsuzawa, H. Protein Eng.,,. Turunen, O., Vuorio, M., Fenel, F., & Leisola, M. Protein Eng.,,. Shirai,T.,Suzuki,A.,Yamane,T.Ashidaa,T.,Kobayashi,T, Hitomi, J., & Ito, S. Protein Eng.,,. Winterhalter, C. & Liebl, W. Appl. Environ. Microbiol.,,. Nelson, K.E., Clayton, R.A., Gill, S.R., Gwinn, M.L., Dodson, R.J., Haft, D.H., Hickey, E.K., Peterson, J.D., Nelson, W.C., Ketchum, K.A., McDonald, L., Utterback, T.R., Malek, J.A., Linher, K.D., Garrett, M.M., Stewart, A.M., Cotton, M.D., Pratt, M.S., Phillips, C.A., Richardson, D., Heidelberg, J., Sutton, G.G., Fleischmann, R.D., Eisen, J.A., White, O., Salzberg, S.L., Smith, H.O., Venter, J.C., & Fraser, C.M. Nature,,. Tsukimura, W., Watanabe, K., Morokuma, C., Ihsanawati, Yatsunami, R., Fukui, T., Kumasaka, T., Tanaka, N., & Nakamura, S. in Biotechnology of Lignocellulose Degradation and Biomass Utilization Sakka, K., Karita, S., Kimura, T., Sakka, M., Matsui, H., Miyake, H. & Tanaka, A. eds., pp., Ito Print Publishing Div., Mie. Ihsanawati, Kumasaka, T., Kaneko, T., Morokuma, C., Yatsunami,R.,Sato,T.,Nakamura,S.,&Tanaka,N. Proteins,,. Manikandan, K., Bhardwaj, A., Gupta, N., Lokanath, N.K., Ghosh, A., Reddy, V.S., & Ramakumar, S. Protein Sci.,,. Mamo, G., Thunnissen, M., Hatti-Kaul, R., & Mattiasson, B. Biochimie,,.

本文／ＹＡ３１８０Ｄ

本文／ＹＡ３１８０Ｄ pp. D- Frontier Science in Amino Acid and Protein Research D- D- N - D- D- D-DAP, EC D-DaaA Ser β- D D- Discovery of D-stereoselective amino acid amidases, and their use in kinetic resolution of amino