title: Molecular cloning and expression of two alpha-amylase genes from Streptococcus bovis 148 in Escherichia coli.
abstract: The alpha-amylase genes of Streptococcus bovis 148 were cloned in Escherichia coli MC1061, using pBR322. The recombinant plasmids were classified into two groups on the basis of their restriction maps. Southern blot analysis did not show homology between the two types of alpha-amylase genes, and the two alpha-amylase genes existed on the chromosomal DNA of S. bovis 148. The enzymatic properties and N-terminal amino acid sequences of the two purified enzymes produced by the cloned E. coli strains were quite different from each other. Particularly, one alpha-amylase (Amy I) was adsorbed on raw corn starch and hydrolyzed raw corn starch, and another (Amy II) was not adsorbed on raw corn starch and did not hydrolyze raw corn starch. Amy I was considered to be the same as the extracellular alpha-amylase of S. bovis 148 in raw starch absorbability, ability to hydrolyze raw corn starch, enzymatic characteristics, N-terminal amino acid sequence, and mode of action on soluble starch. Amy II showed a unique pattern of oligosaccharide production from soluble starch compared with the extracellular alpha-amylase of S. bovis 148. Amy II was suggested to be an intracellular alpha-amylase of S. bovis 148.
authors.affiliation: Department of Agricultural Chemistry, Tokyo University of Agriculture, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Satoh
authors.forename: Y
authors.initials: Y
authors.lastname: Niimura
authors.forename: T
authors.initials: T
authors.lastname: Uchimura
authors.forename: M
authors.initials: M
authors.lastname: Kozaki
authors.forename: K
authors.initials: K
authors.lastname: Komagata
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D013213:Starch / Q000378:metabolism; D016985:Streptococcus bovis / Q000201:enzymology* / Q000235:genetics*; D013696:Temperature; D000516:alpha-Amylases / Q000235:genetics* / Q000378:metabolism
chemical_list: D013213:Starch; D000516:alpha-Amylases