title: Biochemical and genetic characterization of β-1,3 glucanase from a deep subseafloor Laceyella putida.
abstract: A β-1,3-glucanase (LpGluA) of deep subseafloor Laceyella putida JAM FM3001 was purified to homogeneity from culture broth. The molecular mass of the enzyme was around 36 kDa as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). LpGluA hydrolyzed curdlan optimally at pH 4.2 and 80 °C. In spite of the high optimum temperature, LpGluA showed relatively low thermostability, which was stabilized by adding laminarin, xylan, colloidal chitin, pectin, and its related polysaccharides. The gene for LpGluA cloned by using degenerate primers was composed of 1236 bp encoding 411 amino acids. Production of both LpGluA and a chitinase (LpChiA; Shibasaki et al. Appl Microbiol Biotechnol 98, 7845-7853, 2014) was induced by adding N-acetylglucosamine (GluNAc) to a culture medium of strain JAM FM3001. Construction of expression vectors containing the gene for LpGluA and its flanking regions showed the existence of a putative repressor protein.
authors.affiliation: Research & Development Center for Marine Biosciences, Japan Agency for Marine-Earth and Science Technology (JAMSTEC), 2-15 Natsushima, Yokosuka, Kanagawa, 237-0061, Japan. kobayashit@jamstec.go.jp.
authors.forename: Tohru
authors.initials: T
authors.lastname: Kobayashi
authors.affiliation: Research & Development Center for Marine Biosciences, Japan Agency for Marine-Earth and Science Technology (JAMSTEC), 2-15 Natsushima, Yokosuka, Kanagawa, 237-0061, Japan.
authors.forename: Kohsuke
authors.initials: K
authors.lastname: Uchimura
authors.affiliation: Research & Development Center for Marine Biosciences, Japan Agency for Marine-Earth and Science Technology (JAMSTEC), 2-15 Natsushima, Yokosuka, Kanagawa, 237-0061, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Kubota
authors.affiliation: Research & Development Center for Marine Biosciences, Japan Agency for Marine-Earth and Science Technology (JAMSTEC), 2-15 Natsushima, Yokosuka, Kanagawa, 237-0061, Japan.
authors.forename: Takuro
authors.initials: T
authors.lastname: Nunoura
authors.affiliation: Research & Development Center for Marine Biosciences, Japan Agency for Marine-Earth and Science Technology (JAMSTEC), 2-15 Natsushima, Yokosuka, Kanagawa, 237-0061, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Deguchi
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D000595:Amino Acid Sequence; D056449:Bacillales / Q000201:enzymology* / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D019015:Geologic Sediments; D043326:Glucan 1,3-beta-Glucosidase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013696:Temperature; D015533:Transcriptional Activation; D047071:beta-Glucans / Q000378:metabolism
chemical_list: D003470:Culture Media; D047071:beta-Glucans; C038459:curdlan; D043326:Glucan 1,3-beta-Glucosidase; D000117:Acetylglucosamine
keywords: Deep subseafloor; Induction; Laceyella putida; Laminarinase; N-acetylglucosamine; β-1,3-Glucanase