title: A Novel Glycoside Hydrolase Family 5 β-1,3-1,6-Endoglucanase from Saccharophagus degradans 2-40T and Its Transglycosylase Activity.
abstract: In this study, we characterized Gly5M, originating from a marine bacterium, as a novel β-1,3-1,6-endoglucanase in glycoside hydrolase family 5 (GH5) in the Carbohydrate-Active enZyme database. The gly5M gene encodes Gly5M, a newly characterized enzyme from GH5 subfamily 47 (GH5_47) in Saccharophagus degradans 2-40(T) The gly5M gene was cloned and overexpressed in Escherichia coli Through analysis of the enzymatic reaction products by thin-layer chromatography, high-performance liquid chromatography, and matrix-assisted laser desorption ionization-tandem time of flight mass spectrometry, Gly5M was identified as a novel β-1,3-endoglucanase (EC 3.2.1.39) and bacterial β-1,6-glucanase (EC 3.2.1.75) in GH5. The β-1,3-endoglucanase and β-1,6-endoglucanase activities were detected by using laminarin (a β-1,3-glucan with β-1,6-glycosidic linkages derived from brown macroalgae) and pustulan (a β-1,6-glucan derived from fungal cell walls) as the substrates, respectively. This enzyme also showed transglycosylase activity toward β-1,3-oligosaccharides when laminarioligosaccharides were used as the substrates. Since laminarin is the major form of glucan storage in brown macroalgae, Gly5M could be used to produce glucose and laminarioligosaccharides, using brown macroalgae, for industrial purposes.
  
  
  
  In this study, we have discovered a novel β-1,3-1,6-endoglucanase with a unique transglycosylase activity, namely, Gly5M, from a marine bacterium, Saccharophagus degradans 2-40(T) Gly5M was identified as the newly found β-1,3-endoglucanase and bacterial β-1,6-glucanase in GH5. Gly5M is capable of cleaving glycosidic linkages of both β-1,3-glucans and β-1,6-glucans. Gly5M also possesses a transglycosylase activity toward β-1,3-oligosacchrides. Due to the broad specificity of Gly5M, this enzyme can be used to produce glucose or high-value β-1,3- and/or β-1,6-oligosaccharides.
authors.affiliation: Department of Biotechnology, Graduate School, Korea University, Seoul, South Korea.
authors.forename: Damao
authors.initials: D
authors.lastname: Wang
authors.affiliation: Department of Biotechnology, Graduate School, Korea University, Seoul, South Korea.
authors.forename: Do Hyoung
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Graduate School of Analytical Science and Technology and Asia-Pacific Glycomics Reference Site, Chungnam National University, Daejeon, South Korea.
authors.forename: Nari
authors.initials: N
authors.lastname: Seo
authors.affiliation: Department of Biotechnology, Graduate School, Korea University, Seoul, South Korea.
authors.forename: Eun Ju
authors.initials: EJ
authors.lastname: Yun
authors.affiliation: Graduate School of Analytical Science and Technology and Asia-Pacific Glycomics Reference Site, Chungnam National University, Daejeon, South Korea.
authors.forename: Hyun Joo
authors.initials: HJ
authors.lastname: An
authors.affiliation: Department of Food and Nutrition, Chungnam National University, Daejeon, South Korea.
authors.forename: Jae-Han
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Biotechnology, Graduate School, Korea University, Seoul, South Korea khekim@korea.ac.kr.
authors.forename: Kyoung Heon
authors.identifier: http://orcid.org/0000-0003-4600-8668
authors.initials: KH
authors.lastname: Kim
mesh_terms: D002480:Cellulase / Q000235:genetics* / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020563:Gammaproteobacteria / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D005936:Glucans / Q000378:metabolism; D006868:Hydrolysis; D011134:Polysaccharides / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity
chemical_list: D005936:Glucans; D011134:Polysaccharides; C002076:pustulan; C008247:laminaran; D002480:Cellulase