Browse > Article

Cloning of celC, Third Cellulase Gene, from Pectobacterium carotovorum subsp. carotovorum LY34 and its Comparison to Those of Pectobacterium sp.  

LIM WOO JIN (Division of Applied Life Science, Gyeongsang National University)
RYU SUNG KEE (Division of Applied Life Science, Gyeongsang National University)
PARK SANG RYEOL (National Institute of Agricultural Biotechnology, RDA)
KIM MIN KEUN (Division of Applied Life Science, Gyeongsang National University)
AN CHANG LONG (Division of Applied Life Science, Gyeongsang National University)
HONG SU YOUNG (Division of Applied Life Science, Gyeongsang National University)
SHIN EUN CHULE (Division of Applied Life Science, Gyeongsang National University)
LEE JONG YEOUL (National Institute of Agricultural Biotechnology, RDA)
LIM YONG PYO (Department of Horticulture, Chungnam National University)
YUN HAN DAE (Division of Applied Life Science and Research Institute of Life Science, Gyeongsang National University)
Publication Information
Journal of Microbiology and Biotechnology / v.15, no.2, 2005 , pp. 302-309 More about this Journal
Abstract
Phytopathogenic Pectobacterium carotovorum subsp. carotovorum (Pcc) LY34 secretes multiple isozymes of the plant cell wall degrading enzyme endoglucanases. We have cloned a third cel gene encoding CMCase from Pcc LY34. The structural organization of the celC gene (AY188753) consisted of an open reading frame (ORP) of 1,116 bp encoding 371 amino acid residues with a signal peptide of 22 amino acids within the NH$_2$-terminal region of pre-CelC. The predicted amino acid sequence of CelC was similar to that of Peetobaeterium ehrysanthemi Cel8Y (AF282321). The CelC has the conserved region of the glycoside hydrolase family 8. The apparent molecular mass of CelC was calculated to be 39 kDa by CMC-SDS-PAGE. The cellulase­minus mutant of Pee LY34 was as virulent as the wild-type in pathogenicity tests on tubers of potato. The results suggest that the CelC of Pce LY34 is a minor factor for the pathogenesis of soft-rot.
Keywords
Citations & Related Records

Times Cited By Web Of Science : 0  (Related Records In Web of Science)
연도 인용수 순위
  • Reference
1 Guerin, D. M. A., M. B. Lascombe, M. Costtabel, H. Souchon, V. Lamzin, P. Beguin, and P. M. Alzari. 2002. Atomic $(0.94\AA)$ resolution structure of an inverting glycosidase in complex with substrate. J. Mol. Biol. 316: 1061- 1069   DOI   ScienceOn
2 Guiseppi, A., J. L. Aymeric, B. Cami, F. Barras, and N. Creuzet. 1991. Sequence analysis of the cellulase-encoding celY gene of Erwinia chrysanthemi: A possible case of interspccics gene transfer. Gene 106: 109- 114   DOI   ScienceOn
3 Halldorsdottir, S., E. T. Thorolfsdottir, R. Spilliaert, M. Johansson, S. H. Thorbjarnardottir, A. Palsdottir, G. O. Hreggvidsson, J. K. Kristjansson, O. Holst, and G. Eggertsson. 1998. Cloning, sequencing and overexpression of a Rhodothermus marinus gene encoding a thermostable cellulase of glycosyl hydrolase family 12. Appl. Microbiol. Biotechnol. 49: 277- 284   DOI   ScienceOn
4 Henrissat, B. and A. Bairoch. 1993. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem. J. 293: 781- 788   DOI   PUBMED
5 Pissavin, C., J. Robert-Baudouy, and N. Hugouvieux-CottePattat. 1996. Regulation of pelZ, a gene of the pelBC cluster encoding a new pectate lyase in Erwinia chrysanthemi 3937. J. Bacteriol. 178: 7187- 7196   DOI   PUBMED
6 Miller, G. L., R. Blum, W. E. Glennon, and A. L. Burton. 1960. Measurement of carboxylmethylcellulose activity. Anal. Biochem. 2: 127- 132
7 Cho, S. J., S. R. Park, M. K. Kim, W. J. Lim, S. K. Ryu, C. L An, S. Y. Hong, H. Kim, and H. D. Yun. 2002. Cloning of cel8Y gene from Pectobacterium chrysanthemi PY35 and its comparison to eel genes of soft-rot Pectobacterium. Mol. Cells 13: 28- 34   PUBMED
8 Shevchik, V. E., G. Condemine, N. Hugouvieux-CottePattat, and J. Robert-Baudouy. 1996. Characterization of pectin methylesterase B, an outer membrane lipoprotein of Erwinia chrysanthemi 3937. Mol. Microbiol. 19: 445- 466
9 Lymar, E. S., B. Li, and V. Renganathan. 1995. Purification and cellulose-degrading cultures of Phanerochaete chrysosporium. Appl. Environ. Microbiol. 61: 2976- 2980   PUBMED
10 Park, W. Y., S. T. Lim, and H. D. Yun. 1998. Cloning and characterization of a CMCase gene, celB, of Erwinia carotovora subsp. carotovora LY34 and its comparison to celA. Mol. Cells 8: 280- 285   PUBMED
11 Zhou, S. and L. O. Ingram. 2000. Synergistic hydrolysis of carboxymethyl cellulose and acid-swollen cellulose by two endoglucanases (CelZ and CelY) from Erwinia chrysanthemi. Appl. Environ. Microbiol. 67: 6- 14   DOI   ScienceOn
12 Alzari, P. M., H. Souchon, and R. Dominguez. 1996. The crystal structure of endoglucanase CelA, a family 8 glycosyl hydrolase from Clostridium thermocellum. Structure 4: 265- 275   DOI   ScienceOn
13 Lim, S. T., Y. W Park, S. J. Cho, and H. D. Yun. 1997. Phytopathogenicity of Erwinia carotovora subsp. corotovora LY34 and production of CMCase isozymes. Kor. J. Appl. Microbiol. Biotechnol. 25: 468- 476
14 Park, Y. W., S. T. Lim, S. J. Cho, and H. D. Yun. 1997. Characterization of Erwinia carotovora subsp. carotovora LY34 $endo-1,4-\beta-glucanase$ genes and rapid identification of their gene products. Biochem. Biophys. Res. Cummun. 241: 636- 641   DOI   PUBMED   ScienceOn
15 Saarilahti, H. T., B. Henrissat, and E. T. Paiva. 1990. CelS: A novel endoglucanase identified from Erwinia carotovora subsp. carotovora. Gene 90: 9- 14   DOI   ScienceOn
16 Nakamura, K., N. Misawa, and K. Kitamura. 1986. Sequence of a cellulase gene of Cellulomonas uda CB4. J. Biotechnol. 4: 247- 254   DOI   ScienceOn
17 Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254   DOI   PUBMED   ScienceOn
18 Dale, B. E. 1999. Biobased industrial products: Bioprocess engineering when cost really counts. Biotechnol. Prog. 15: 775-776   DOI   PUBMED   ScienceOn
19 Mae, A. R., R. Heikinheimo, and E. T. Palva. 1995. Structure and regulation of the Erwinia carotovora SCC3193 cellulase gene celV1 and the role of cellulase in phytopathogenicity. Mol. Gen. Genet. 247: 17- 26   DOI   ScienceOn
20 Sambrook, J. and D. W. Russel. 2001. Molecular Cloning: A Laboratory Manual, 3rd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, U.S.A
21 Hong, I. P, H. K. Jang, S. Y. Lee, and S. G. Choi. 2003. Cloning and characterization of a bifunctional cellulasechitosanasc gene from Bacillus licheniformis NBL420. J. Microbiol. Biotechnol. 13: 35- 42
22 Park, S. R., M. K. Kim, J. O. Kim, D. W. Bae, S. J. Cho, Y. U. Cho, and H. D. Yun, 2000. Characterization of Erwinia chrysanthemi PY35 cel and pel genes existing in tandem and rapid identification of their gene products. Biochem. Biophys. Res. Commun. 268: 420- 425   DOI   ScienceOn
23 Kim, K. C., S. S. Yoo, Y. A Oh, and S. J. Kim. 2003. Isolation and characteristics of Trichoderma harzianum FJ1 producing cellulases and xylanase. J. Microbiol. Biotechnol. 13: 1-8
24 Kim, K. J., Y. J. Yang, and J. K. Kim. 2002. Production of $\alpha$-glucosidase inhibitor by $\beta$-glucosidase inhibitor-producing Bacillus lentlmorbus B-6. J. Microbiol. Biotechnol. 12: 895- 900
25 Ryu, S. K., S. R. Park, W. J. Lim, M. K. Kim, C. L. An, S. J. Cho, Y. W. Park, B. K. Kim, H. Kim, and H. D. Yun. 2001. Cloning and characterization of the cel9A gene of marine bacterium Pseudomonas sp. SK38 causing green spot rot of Porphyra dentata. Appl. Microbiol. Biotechnol. 57: 138-145   DOI   ScienceOn
26 Beguin, P. 1990. Molecular biology of cellulose degradation. Annu. Rev. Microbiol. 44: 219- 248   DOI   PUBMED   ScienceOn
27 Cooper, V. J. C. and G. P. C. Salmond. 1993. Molecular analysis of the major cellulase (CeIV) of Erwinia carotovora: Evidence for an evolutionary mix-and-match of enzyme domains. Mol. Gen. Genet. 241: 341- 350   PUBMED
28 Johnvesly, B., S. Virupakshi, G. N. P. Ramalingam, and G. R. Naik. 2002. Cellulase-free thermostable alkaline xylanase from thermophilic and alkalophilic Bacillus sp. JB-99. J. Microbiol. Biotechnol. 12: 153- 156
29 Aymeric, J. L., A. Guiseppi, M. C. Pascal, and M. Chippaux. 1988. Mapping and regulation of the cel genes in Erwinia chrysanthemi. Mol. Gen. Genet. 211: 95- 101   DOI   ScienceOn
30 Boyer, M. H., B. Cami, A. Koutoujansky, J. P. Charnbost, C. Frixon, and J. Cattaneo. 1987. Isolation of the gene encoding the major endoglucanase of Erwinia chrysanthemi; homology between celZ genes of two strains of Erwinia chrysanthemi. FEMS Microbiol. Lett. 41: 351- 356   DOI   ScienceOn