title: A neopullulanase-type alpha-amylase gene from Thermoactinomyces vulgaris R-47.
abstract: Shimizu et al. and ourselves have reported some enzymatic properties of an alpha-amylase from Thermoactinomyces vulgaris R-47 that hydrolyzed pullulan to produce panose [M. Shimizu et al., Agric. Biol. Chem., 42, 1681-1688 (1978); Y. Sakano et al., Agric. Biol. Chem., 46, 1121-1129 (1982)]. In this study, we cloned a gene for an alpha-amylase, which was different from the one mentioned above but also hydrolyzed pullulan to produce panose, from T. vulgaris R-47, and analyzed the entire primary structure of the gene. We designated the previously reported enzyme as T. vulgaris alpha-amylase I (TVA I), and this novel enzyme as T. vulgaris alpha-amylase II (TVA II). The nucleotide sequence had an open reading frame of 1755 base pairs corresponding to a protein of 585 amino acid residues. Although this novel alpha-amylase, TVA II, hydrolyzed both pullulan and starch, the ratio of pullulan-hydrolyzing activity to starch-hydrolyzing activity of the enzyme was higher than that of TVA I, and the primary structure of the enzyme resembled neopullulanase, which scarcely hydrolyzed starch, rather than that of TVA I.
authors.affiliation: Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Tonozuka
authors.forename: M
authors.initials: M
authors.lastname: Ohtsuka
authors.forename: S
authors.initials: S
authors.lastname: Mogi
authors.forename: H
authors.initials: H
authors.lastname: Sakai
authors.forename: T
authors.initials: T
authors.lastname: Ohta
authors.forename: Y
authors.initials: Y
authors.lastname: Sakano
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D005798:Genes, Bacterial; D005936:Glucans / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry*; D008848:Micromonosporaceae / Q000201:enzymology*; D008969:Molecular Sequence Data; D010957:Plasmids; D014158:Transcription, Genetic; D000516:alpha-Amylases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D004269:DNA, Bacterial; D005936:Glucans; C009109:pullulan; D006026:Glycoside Hydrolases; D000516:alpha-Amylases; C058484:neopullulanase