title: The fucosidase-pool of Emticicia oligotrophica: Biochemical characterization and transfucosylation potential.
abstract: Three novel bacterial α-l-fucosidases, which cleave terminal fucosyl residues from glycoconjugates are reported in this work. Originating from the recently discovered bacterium Emticicia oligotrophica, recombinant fucosidase isoforms designated as Eo0918, Eo3066 and Eo3812 were shown to have the highest activity between pH 6.0 and 7.0 and temperature optima between 30 and 45°C. All enzymes catalyzed the hydrolysis of the model substrate pNP-α-l-fucose and revealed significantly different regiospecificities towards fucose-containing oligosaccharides: Eo0918 liberated exclusively α1,6-linked fucose and Eo3812 released only α1,3-fucosyl residues, whereas Eo3066 showed broader substrate promiscuity. The enzymatic activity of Eo0918 and Eo3812 increased upon the addition of Ca(2+), Mn(2+) and Zn(2+) ions, whereas the activity of Eo3066 was significantly decreased in the presence of these metal ions. In addition, Eo0918 also catalyzed the transfer of fucose from pNP-α-l-fucose to the 7-hydroxyl group of 4-methylumbelliferone with up to 15% transglycosylation yield. Facile recombinant expression in E. coli, distinct substrate specificities and the transglycosylation ability of Eo0918 presented herein make these newly discovered fucosidases valuable candidates for bioanalytical and biotechnological applications.
authors.affiliation: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors.forename: Si
authors.initials: S
authors.lastname: Liu
authors.affiliation: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors.forename: Anna
authors.initials: A
authors.lastname: Kulinich
authors.affiliation: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors.forename: Zhi P
authors.initials: ZP
authors.lastname: Cai
authors.affiliation: Department of Plant Pathology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors.forename: Hong Y
authors.initials: HY
authors.lastname: Ma
authors.affiliation: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors.forename: Ya M
authors.initials: YM
authors.lastname: Du
authors.affiliation: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors.forename: Yong M
authors.initials: YM
authors.lastname: Lv
authors.affiliation: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors.forename: Josef
authors.initials: J
authors.lastname: Voglmeir
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D041963:Bacteroidetes / Q000737:chemistry / Q000201:enzymology*; D055162:Biocatalysis; D002118:Calcium / Q000737:chemistry; D002413:Cations, Divalent; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005643:Fucose / Q000737:chemistry* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D006923:Hymecromone / Q000737:chemistry; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008345:Manganese / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity; D013696:Temperature; D015032:Zinc / Q000737:chemistry; D005644:alpha-L-Fucosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D002413:Cations, Divalent; D007527:Isoenzymes; D011994:Recombinant Proteins; D005643:Fucose; D006923:Hymecromone; D008345:Manganese; D005644:alpha-L-Fucosidase; D015032:Zinc; D002118:Calcium
keywords: Emticicia oligotrophica; bacterial glycosidases; fucose-containing oligosaccharide; fucosidase; transfucosylation