Characterization of a PA14 domain-containing galactofuranose-specific β-d-galactofuranosidase from Streptomyces sp.
As a constituent of polysaccharides and glycoconjugates, β-d-galactofuranose (Galf) exists in several pathogenic microorganisms. Although we recently identified a β-d-galactofuranosidase (Galf-ase) gene, ORF1110, in the Streptomyces strain JHA19, very little is known about the Galf-ase gene. Here, we characterized a strain, named JHA26, in the culture supernatant of which exhibited Galf-ase activity for 4-nitrophenyl β-d-galactofuranoside (pNP-β-d-Galf) as a substrate. Draft genome sequencing of the JHA26 strain revealed a putative gene, termed ORF0643, that encodes Galf-ase containing a PA14 domain, which is thought to function in substrate recognition. The recombinant protein expressed in Escherichia coli showed the Galf-specific Galf-ase activity and also released galactose residue of the polysaccharide galactomannan prepared from Aspergillus fumigatus, suggesting that this enzyme is an exo-type Galf-ase. BLAST searches using the amino acid sequences of ORF0643 and ORF1110 Galf-ases revealed two types of Galf-ases in Actinobacteria, suggesting that Galf-specific Galf-ases may exhibit discrete substrate specificities.
a Faculty of Agriculture, Department of Bioscience and Biotechnology , Kyushu University , Fukuoka , Japan.
Emiko
E
Matsunaga
a Faculty of Agriculture, Department of Bioscience and Biotechnology , Kyushu University , Fukuoka , Japan.
Yujiro
Y
Higuchi
a Faculty of Agriculture, Department of Bioscience and Biotechnology , Kyushu University , Fukuoka , Japan.
Kazuki
K
Mori
a Faculty of Agriculture, Department of Bioscience and Biotechnology , Kyushu University , Fukuoka , Japan.
Nao
N
Yairo
a Faculty of Agriculture, Department of Bioscience and Biotechnology , Kyushu University , Fukuoka , Japan.
Saki
S
Toyota
b Faculty of Biotechnology and Life Science, Department of Applied Microbial Technology , Sojo University , Kumamoto , Japan.
Takuji
T
Oka
a Faculty of Agriculture, Department of Bioscience and Biotechnology , Kyushu University , Fukuoka , Japan.
Kosuke
K
Tashiro
a Faculty of Agriculture, Department of Bioscience and Biotechnology , Kyushu University , Fukuoka , Japan.
Kaoru
K
Takegawa
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005690:Galactose / Q000031:analogs & derivatives* / Q000378:metabolism; D005697:Galactosides / Q000737:chemistry* / Q000378:metabolism; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008351:Mannans / Q000737:chemistry* / Q000378:metabolism; D010802:Phylogeny; D000072417:Protein Domains; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000737:chemistry* / Q000145:classification / Q000201:enzymology; D013379:Substrate Specificity
D001426:Bacterial Proteins; D005697:Galactosides; D008351:Mannans; D011994:Recombinant Proteins; C060549:4-nitrophenylgalactofuranoside; C012990:galactomannan; D006026:Glycoside Hydrolases; C023112:exo-beta-D-galactofuranosidase; D005690:Galactose
Streptomyces species; galactomannan; α-l-arabinofuranosidase; β-d-galactofuranosidase