title: Sequential processing of mannose-containing glycans by two α-mannosidases from Solitalea canadensis.
abstract: Two putative α-mannosidase genes isolated from the rather unexplored soil bacterium Solitalea canadensis were cloned and biochemically characterised. Both recombinant enzymes were highly selective in releasing α-linked mannose but no other sugars. The α-mannosidases were designated Sca2/3Man2693 and Sca6Man4191, and showed the following biochemical properties: the temperature optimum for both enzymes was 37 °C, and their pH optima lay at 5.0 and 5.5, respectively. The activity of Sca2/3Man2693 was found to be dependent on Ca(2+) ions, whereas Cu(2+) and Zn(2+) ions almost completely inhibited both α-mannosidases. Specificity screens with various substrates revealed that Sca2/3Man2693 could release both α1-2- and α1-3-linked mannose, whereas Sca6Man4191 only released α1-6-linked mannose. The combined enzymatic action of both recombinant α-mannosidases allowed the sequential degradation of high-mannose-type N-glycans. The facile expression and purification procedures in combination with strict substrate specificities make α-mannosidases from S. canadensis promising candidates for bioanalytical applications.
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors: Fang F
authors: FF
authors: Liu
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors: Anna
authors: A
authors: Kulinich
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.
authors: Ya M
authors: YM
authors: Du
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China. liu.lichen@qlyco.com.|Qlyco Ltd., Nanjing, People's Republic of China. liu.lichen@qlyco.com.
authors: Li
authors: L
authors: Liu
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China. josef.voglmeir@njau.edu.cn.
authors: Josef
authors: J
authors: Voglmeir
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D041963:Bacteroidetes / Q000201:enzymology* / Q000235:genetics; D008358:Mannose / Q000737:chemistry*; D013379:Substrate Specificity; D043323:alpha-Mannosidase / Q000737:chemistry* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D043323:alpha-Mannosidase; D008358:Mannose
keywords: Bacterial glycosidases; Mannose-containing oligosaccharide; Mannosidase; N-glycans; Oligomannose; Solitalea canadensis