title: Cloning of the cel8Y gene from Pectobacterium chrysanthemi PY35 and its comparison to cel genes of soft-rot Pectobacterium.
abstract: The phytopathogenic Pectobacterium chrysanthemi (Pch) PY35 secretes multiple isozymes of plant cell wall degrading enzyme cellulases. We cloned a second cel gene that encodes cellulase in Pch PY35. The inserted 2 kb fragment was subcloned in order to geneate pPY710 (cel8Y). The structural organization of the cel8Y gene consists of an open reading frame (ORF) of 999 bp that encodes 332 amino acid residues with a signal peptide of 23 amino acids. The predicted amino acid sequence of Cel8Y was very similar to that of Cellulomonas uda, but completely different from that of the Cel5Z of Pch PY35. It belonged to the glycoside hydrolase family 8, based on amino acid sequence similarities in contrast to Cel5Z of Pch PY35, which was confirmed as family 5. Cel8Y was not closely related to the known cellulases of Pectobacterium. It had the conserved region of the glycoside hydrolase family 8, ASDGDVLIAWALLKAGNKW. The apparent molecular mass of the Cel8Y protein was calculated to be approximately 34 kDa by a carboxymethylcellulosesodium dodecyl sulfate-polyacrylamide gel electrophoresis (CMC-SDS-PAGE). The Cel8Y had a calculated pl of 6.49. It was optimally active at pH 7 with an approximate optimal temperature around 40 degrees C. The cellulase activity of Cel8Y was lower than that of Cel5Z.
authors.affiliation: Division of Applied Life Science, Gyeongsang National University, Chinju, Korea.
authors.forename: Soo Jeong
authors.initials: SJ
authors.lastname: Cho
authors.forename: Sang Ryeol
authors.initials: SR
authors.lastname: Par
authors.forename: Min Keun
authors.initials: MK
authors.lastname: Kim
authors.forename: Woo Jin
authors.initials: WJ
authors.lastname: Lim
authors.forename: Sung Kee
authors.initials: SK
authors.lastname: Ryu
authors.forename: Chang Long
authors.initials: CL
authors.lastname: An
authors.forename: Su Young
authors.initials: SY
authors.lastname: Hong
authors.forename: Hoon
authors.initials: H
authors.lastname: Kim
authors.forename: Yong Un
authors.initials: YU
authors.lastname: Cho
authors.forename: Han Dae
authors.initials: HD
authors.lastname: Yun
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002480:Cellulase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004269:DNA, Bacterial / Q000235:genetics; D016974:Dickeya chrysanthemi / Q000201:enzymology* / Q000235:genetics*; D019143:Evolution, Molecular; D005798:Genes, Bacterial; D006026:Glycoside Hydrolases / Q000235:genetics; D007526:Isoelectric Point; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D010802:Phylogeny; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D007527:Isoenzymes; D006026:Glycoside Hydrolases; D002480:Cellulase; C449655:cel5Z protein, Erwinia chrysanthemi