研究成果報告書(基金分)

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2 N- GlcNAc β-1,4 GlcNAc GlcNAc DNA Neisseriaceae Chitiniphilus shinanonensis SAY3 T SAY3 GlcNAc C. shinanonensis SAY3 DNA SAY3 ORF pcold, pet GlcNAc HPLC pmo130 SAY3 SAY3 15

3 SAY3 DNA GC 67.6% 4.16 x kbp contig 86 3,278 ORF 15 chia-chio 34 chi1-chi34 SAY3 49 ChiA ChiB ChiC ChiD ChiE ChiF ChiG ChiH ChiI ChiJ ChiK ChiL ChiM ChiN ChiO Chi1 Chi2 Chi3 Chi4 Chi5 Chi6 Chi7 Chi8 Chi9 Chi10 GH18 GH 20 GH 19 GH 19 LytTR ChiC ChiC Chi11 Chi12 Chi13 Chi14 Chi15 Chi16 Chi17 Chi18 Chi19 Chi20 Chi21 Chi22 Chi23 Chi24 Chi25 Chi26 Chi27 Chi28 Chi29 Chi30 Chi31 Chi32 Chi33 Chi34 GH 16 CBM 6 GH 19 FGE-sulfatase ChBD3 Esterase GH 16 2 SAY3 SAY ChiG 2 LytTR 18 GH18 ChiG GlcNAc 2 6 GlcNAc GlcNAc HPLC 5 5 β α ChiG GlcNAc retention ChiG GlcNAc Standard (GlcNAc) 1-6 Initial reaction products from (GlcNAc) 6 59:41 1α 1β 25:75 1β 1α 62:38 63:37 64:36 3α 2α 4α 2β 3β 4β 25:75 2β 2α 31:69 3β 3α 45:54 4β 4α retention time (min) 67:33 5α 5β column BEH amide column solvent 70% acetonitrile UV detection O.D :33 6α 6β 37:63 73:27 6α 5β 6β 5α ChiG GlcNAc6 ChiJ, ChiK GH18 GlcNAc processive ChiL GH18 GlcNAc3 2 ChiL

4 Chi33 SAY3 pmo130 SAY3 DNA SAY3 N- GlcNAcase GH20 chii 2% GlcNAcase > Kazuaki Sato, Yuichi Kato, Goro Taguchi, Masahiro Nogawa, Akira Yokota, and Makoto Shimosaka: Chitiniphilus shinanonensis gen. nov., sp. nov., a novel chitin-degrading bacterium belonging to Betaproteobacteria; J. Gen. Appl. Microbiol. 55, , (2009). Hamad M. A., Zajdowicz S. L., Holmes R. K., and Voskuil M. I.: An allelic exchange system for compliant genetic manipulation of the select agents Burkholderia pseudomallei and Burkholderia mallei; Gene, 430 (1), (2009). Lanxiang Huang, Ewelina Garbulewska, Kazuaki Sato, Yuichi Kato, Masahiro Nogawa, Goro Taguchi, and Makoto Shimosaka: Isolation of genes coding for chitin-degrading enzymes in the novel chitinolytic bacterium, Chitiniphilus shinanonensis, and characterization of a gene coding for a family 19 chitinase; J. Biosci. Bioeng. 113(3), , (2012). chii C. shinanonensis SAY3 Chitiniphilus shinanonensis N- 30(4), 52-59, (2013).

5 Lanxiang Huang, Arisa Shizume, Masahiro Nogawa, Goro Taguchi, and Makoto Shimosaka: Heterologous expression and functional characterization of a novel chitinase from the chitinolytic bacterium, Chitiniphilus shinanonensis; Biosci. Biotechnol. Biochem. 76(3), , (2012). DOI: /bbb Chitiniphilus shinanonensis ChiJ, ChiK Chitiniphilus shinanonensis (Moe Nakano), Jiayi Yu, Lanxiang Huang, Masahiro Nogawa, Goro Taguchi, Makoto Shimosaka: Analysis of genes coding for chitinolytic enzymes in the bacterium, Chitiniphilus shinanonensis chig, chij, and chik coding for unique chitin-degrading enzymes ;the 10th International Conference of the Asian Pacific Chitin Chitosan Symposium, ( ) (Norie Sonoda), Jiayi Yu, Arisa Shizume, Masahiro Nogawa, Goro Taguchi, Makoto Shimosaka Analysis of genes coding for chitinolytic enzymes in the bacterium, Chitiniphilus shinanonensis Development of gene disruption by an allelic exchange system ; the 10th International Conference of the Asian Pacific Chitin Chitosan Symposium, ( ) (Makoto Shimosaka), Lanxiang Huang, Norie Sonoda, Moe Nakano, Masahiro Nogawa, Goro Taguchi: Isolation and characterization of genes encoding chitin-degrading enzymes in the novel chitinolytic bacterium, Chitiniphilus shinanonensis; the 10th International Conference of the Asian Pacific Chitin Chitosan Symposium, () ( ) Chitiniphilus shinanonensis ChiG Chitiniphilus shinanonensis ChiL : Chitiniphilus shinanonensis

6 (1) SHIMOSAKA, Makoto (2) TAGUCHI, Goro

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