Nucleotide and deduced amino acid sequences of mutanase-like genes from Paenibacillus isolates: proposal of a new family of glycoside hydrolases.
Three mutanase (alpha-1,3-glucanase)-producing microorganisms isolated from soil samples were identified as a relatives of Paenibacillus. A mutanase was purified to homogeneity from cultures of each, and the molecular masses of the purified enzymes were approximately 132, 141, and 141kDa, respectively. The corresponding three genes for mutanases were cloned by PCR using primers designed from each N-terminal amino acid sequence. Another mutanase-like gene from one strain was also cloned by PCR using primers designed from conserved amino acid sequences among known mutanases. Consequently, four mutanase-like genes were sequenced. The genes contained long open reading frames of 3411 to 3915bp encoding 1136 to 1304 amino acids. The deduced amino acid sequences of the mutanases showed relatively high similarity to those of a mutanase (E16590) from Bacillus sp. RM1 with 46.9% to 73.2% identity and an alpha-1,3-glucanase (AB248056) from Bacillus circulans KA-304 with 46.7% to 70.4% identity. Phylogenetic analysis based on the amino acid sequences of the enzymes showed bacterial mutanases form a new family between fungal mutanases (GH family 71) and Streptomycetes mycodextranases (GH family 87).
Hakamada
Yoshihiro
Y
Tochigi Research Laboratories of Kao Corporation, Haga, Tochigi, Japan.
Sumitomo
Nobuyuki
N
Ogawa
Akinori
A
Kawano
Takako
T
Saeki
Katsuhisa
K
Ozaki
Katsuya
K
Ito
Susumu
S
Kobayashi
Tohru
T
D000595:Amino Acid Sequence; D001419:Bacteria / Q000145:classification / Q000201:enzymology*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000145:classification* / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid
D006026:Glycoside Hydrolases; C021780:exo-1,3-alpha-glucanase