title: Characterization of α-1,3-glucanase isozyme from Paenibacillus glycanilyticus FH11 in a new subgroup of family 87 α-1,3-glucanase.
abstract: Two α-1,3-glucanase isozymes, designated as α-1,3-glucanase 1 (Agl-FH1) and α-1,3-glucanase 2 (Agl-FH2), were purified from the culture medium of Paenibacillus glycanilyticus FH11. Agl-FH1 and Agl-FH2 exhibited similar characteristics such as optimal pH, pH stability, optimal temperature, thermostability, and molecular masses on SDS-PAGE. However, their hydrolysis products of α-1,3-glucan varied somewhat. Agl-FH1 hydrolyzed α-1,3-glucan into a mixture of maltotriose and maltotetraose, and maltotetraose was the major hydrolysis product of Agl-FH2. N-terminal amino acid sequence analysis and LC-MS/MS analysis of trypsin digested fragments revealed several differences between the amino acid sequences of Agl-FH1 and Agl-FH2. Genes of Agl-FH1 and Agl-FH2 were subcloned into an expression plasmid, and both enzymes were successfully expressed in Escherichia coli. The recombinant Agl-FH1 and Agl-FH2 exhibited the same enzymatic properties as those of each wild-type enzyme, and both of the recombinants showed the activity on the protoplast formation of Schizophyllum commune mycelia. A great diversity was detected in the C-terminal region of family 87 α-1,3-glucanases. Compared with Agl-FH2 which is highly sequence-related to the known α-1,3-glucanases, the C-terminal region of Agl-FH1 has only slight similarity to them (approximately 20% identity). Our analysis revealed that Agl-FH1 was the first member of a new subgroup of family 87 α-1,3-glucanases.
authors.affiliation: Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Wasana
authors.initials: W
authors.lastname: Suyotha
authors.affiliation: Department of Biochemical Engineering, Graduate School of Sciences and Engineering, Yamagata University, Jonan, Yonezawa, Yamagata 992-8510, Japan.
authors.forename: Shigekazu
authors.initials: S
authors.lastname: Yano
authors.affiliation: Department of Bioscience, Faculty of Biotechnology, Fukui Prefectural University, Eiheiji-cho, Yoshida-gun, Fukui 910-1195, Japan.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Itoh
authors.affiliation: Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Fujimoto
authors.affiliation: Department of Bioscience, Faculty of Biotechnology, Fukui Prefectural University, Eiheiji-cho, Yoshida-gun, Fukui 910-1195, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Hibi
authors.affiliation: Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Tachiki
authors.affiliation: Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University, Kusatsu, Shiga 525-8577, Japan. Electronic address: wakayama@sk.ritsumei.ac.jp.
authors.forename: Mamoru
authors.initials: M
authors.lastname: Wakayama
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005936:Glucans / Q000737:chemistry* / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D008320:Maltose / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D025282:Mycelium / Q000187:drug effects / Q000254:growth & development; D056507:Paenibacillus / Q000737:chemistry* / Q000201:enzymology; D017434:Protein Structure, Tertiary; D011523:Protoplasts / Q000187:drug effects; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D012567:Schizophyllum / Q000187:drug effects / Q000254:growth & development; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D005936:Glucans; D007527:Isoenzymes; D011994:Recombinant Proteins; D008320:Maltose; C045788:alpha-1,3-glucan; C009819:maltotetraose; D006026:Glycoside Hydrolases; C021780:exo-1,3-alpha-glucanase
keywords: Characterization; Isozyme; Paenibacillus glycanilyticus FH11; Protoplasts formation; α-1,3-Glucanase