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Start Over Author "Saki Shinozuka" Publisher public library of science (plos)
2 results on '"Saki Shinozuka"'

1. Identification and Characterization of a Novel Galactofuranose-Specific β-D-Galactofuranosidase from Streptomyces Species.

Author

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

Subjects
Medicine, Science
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.

Published
2015
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2. Identification and Characterization of a Novel Galactofuranose-Specific β-D-Galactofuranosidase from Streptomyces Species

Author

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

Subjects
animal structures, Glycoside Hydrolases, Molecular Sequence Data, lcsh:Medicine, Sequence alignment, medicine.disease_cause, Genome, Streptomyces, Microbiology, Substrate Specificity, Mannans, Open Reading Frames, Bacterial Proteins, RNA, Ribosomal, 16S, Hydrolase, medicine, Escherichia coli, Amino Acid Sequence, Cloning, Molecular, lcsh:Science, Peptide sequence, Gene, Phylogeny, Multidisciplinary, biology, Sequence Homology, Amino Acid, Aspergillus fumigatus, lcsh:R, Galactose, High-Throughput Nucleotide Sequencing, Ribosomal RNA, Hydrogen-Ion Concentration, biology.organism_classification, Recombinant Proteins, Kinetics, Biochemistry, lcsh:Q, Glycoconjugates, Sequence Alignment, Genome, Bacterial, Research Article
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.

Published
2015

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