Identification and characterization of a novel galactofuranose-specific β-D-galactofuranosidase from Streptomyces species

Emiko Matsunaga, Yujiro Higuchi, Kazuki Mori, Nao Yairo, Takuji Oka, Saki Shinozuka, Kosuke Tashiro, Minoru Izumi, Satoru Kuhara, Kaoru Takegawa

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

β-D-galactofuranose (Galf) is a component of polysaccharides and glycoconjugates and its transferase has been well analyzed. However, no β-D-galactofuranosidase (Galf-ase) gene has been identified in any organism. To search for a Galf-ase gene we screened soil samples and discovered a strain, identified as a Streptomyces species by the 16S ribosomal RNA gene analysis, that exhibits Galf-ase activity for 4-nitrophenyl β-D-galactofuranoside (pNP-β-D-Galf) in culture supernatants. By draft genome sequencing of the strain, named JHA19, we found four candidate genes encoding Galf-ases. Using recombinant proteins expressed in Escherichia coli, we found that three out of four candidates displayed the activity of not only Galf-ase but also α-L-arabinofuranosidase (Araf-ase), whereas the other one showed only the Galf-ase activity. This novel Galf-specific hydrolase is encoded by ORF1110 and has an optimum pH of 5.5 and a Km of 4.4 mM for the substrate pNP-β-D-Galf. In addition, this enzyme was able to release galactose residue from galactomannan prepared from the filamentous fungus Aspergillus fumigatus, suggesting that natural polysaccharides could be also substrates. By the BLAST search using the amino acid sequence of ORF1110 Galf-ase, we found that there are homolog genes in both prokaryotes and eukaryotes, indicating that Galf-specific Galf-ases widely exist in microorganisms.

Original languageEnglish
Article numbere0137230
JournalPloS one
Volume10
Issue number9
DOIs
Publication statusPublished - Sep 4 2015

Fingerprint

Streptomyces
Genes
Polysaccharides
genes
16S Ribosomal RNA
polysaccharides
Glycoconjugates
Aspergillus fumigatus
alpha-N-arabinofuranosidase
Hydrolases
Transferases
Eukaryota
Galactose
rRNA Genes
Recombinant Proteins
glycoconjugates
Gene encoding
Aspergillus
Amino Acid Sequence
Substrates

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Identification and characterization of a novel galactofuranose-specific β-D-galactofuranosidase from Streptomyces species. / Matsunaga, Emiko; Higuchi, Yujiro; Mori, Kazuki; Yairo, Nao; Oka, Takuji; Shinozuka, Saki; Tashiro, Kosuke; Izumi, Minoru; Kuhara, Satoru; Takegawa, Kaoru.

In: PloS one, Vol. 10, No. 9, e0137230, 04.09.2015.

Research output: Contribution to journalArticle

Matsunaga, Emiko ; Higuchi, Yujiro ; Mori, Kazuki ; Yairo, Nao ; Oka, Takuji ; Shinozuka, Saki ; Tashiro, Kosuke ; Izumi, Minoru ; Kuhara, Satoru ; Takegawa, Kaoru. / Identification and characterization of a novel galactofuranose-specific β-D-galactofuranosidase from Streptomyces species. In: PloS one. 2015 ; Vol. 10, No. 9.
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