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

PloS One
Emiko MatsunagaKaoru Takegawa

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-a...Continue Reading

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Citations

Mar 21, 2017·Bioscience, Biotechnology, and Biochemistry·Emiko MatsunagaKaoru Takegawa
Jan 16, 2018·Bioscience, Biotechnology, and Biochemistry·Takuji Oka
Feb 29, 2020·The Journal of Biological Chemistry·Lin ShenChristophe Mariller
May 20, 2020·International Journal of Molecular Sciences·Mateja SeničarRichard Daniellou
Oct 5, 2020·Journal of Bioscience and Bioengineering·Emiko MatsunagaKaoru Takegawa
Jun 8, 2019·Carbohydrate Research·Mateja SeničarRichard Daniellou
Jun 27, 2021·Metabolic Engineering·Diana Chinyere AnyaoguUffe Hasbro Mortensen

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Datasets Mentioned

BETA
ORF2125
ORF2812

Methods Mentioned

BETA
PCR
whole-genome shotgun sequencing
ELISA

Software Mentioned

MetaVue
CLUSTAL
BLAST
Newbler
CLUSTAL W
Pfam
Glimmer

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