title: Cloning and sequencing analysis of alginate lyase genes from the marine bacterium Vibrio sp. O2.
abstract: We isolated a new marine bacteria, which displayed alginate-depolymerizing activity in plate assays, from seawater in Mihonoseki Harbor, Japan. Analysis of the 16S ribosomal RNA gene sequence of one of the isolates proved that this alginate-depolymerizing bacterium belonged to the genus Vibrio and it was named Vibrio sp. O2. The alginate lyase genes of Vibrio sp. O2 were cloned and expressed in Escherichia coli. Two alginate lyase-producing clones, pVOA-A4 and pVOA-B5, were obtained. The alginate lyase gene alyVOA from pVOA-A4 was composed of an 858-bp open reading frame (ORF) encoding 285 amino acid residues, while alyVOB from pVOA-B5 was composed of an 828-bp ORF encoding 275 amino acid residues. The degree of identity between the deduced amino acid sequences of AlyVOA or AlyVOB and Photobacterium sp. ATCC43367 alginate poly(ManA)lyase AlxM was 92.3% or 32.6%, respectively. Alginate lyase consensus regions corresponding to the sequences YFKAGXYXQ and RXELR were observed in all three of these sequences. AlyVOA and AlyVOB both degraded polymannuronate in plate assays and were therefore confirmed to be poly(beta-D-mannuronate)lyases.
authors.affiliation: Marine Products Kimuraya Co Ltd, 3307 Watari, Sakaiminato, Tottori, Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Kawamoto
authors.forename: Akio
authors.initials: A
authors.lastname: Horibe
authors.forename: Yasunari
authors.initials: Y
authors.lastname: Miki
authors.forename: Takayuki
authors.initials: T
authors.lastname: Kimura
authors.forename: Katsunori
authors.initials: K
authors.lastname: Tanaka
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Makoto
authors.initials: M
authors.lastname: Kawamukai
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Matsuda
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014733:Vibrio / Q000201:enzymology* / Q000235:genetics*
chemical_list: D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase