title: 2-Keto-D-gluconate-yielding membrane-bound D-glucose dehydrogenase from Arthrobacter globiformis C224: purification and characterization.
abstract: Glucose dehydrogenase (GlcDH) is the rate-limiting catalyst for microbial conversion of glucose to the important organic acid 2-ketogluconic acid (2KGlcA). In this study, a D-glucose dehydrogenase was purified from the industrial 2KGlcA producer Arthrobacter globiformis C224. After four purification steps, the GlcDH was successfully purified over 180 folds and specific activity of 88.1 U/mg. A single protein band of 87 kDa was detected by SDS-PAGE. The purified GlcDH had the broad substrate specificity with the Km values for D-glucose, D-xylose, D-galactose and maltose of 0.21 mM, 0.34 mM, 0.46 mM and 0.59 mM, respectively. The kinetic studies proved that A. globiformis GlcDH followed the ping-pong kinetic mechanism. The GlcDH showed an optimum catalytic activity at pH 5.0 and 45 °C with the stable activity at temperature of 20-40 °C and pH of 6.0-7.0. Organic solvents, metal ions or EDTA could significantly influence the GlcDH activity to different degrees.
authors.affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China. liujingze@mail.hebtu.edu.cn.
authors.forename: Qing
authors.initials: Q
authors.lastname: Xue
authors.affiliation: Parchn Sodium Isovitamin C Co. Ltd, Dexing 334221, China. weizhuan8856@163.com.
authors.forename: Zhuan
authors.initials: Z
authors.lastname: Wei
authors.affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China. sunwenjing1919@163.com.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Sun
authors.affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China. fengjiecui@163.com.
authors.forename: Fengjie
authors.initials: F
authors.lastname: Cui
authors.affiliation: Parchn Sodium Isovitamin C Co. Ltd, Dexing 334221, China. ysl@parchn.com.
authors.forename: Silian
authors.initials: S
authors.lastname: Yu
authors.affiliation: Parchn Sodium Isovitamin C Co. Ltd, Dexing 334221, China. zhouqiang@parchn.com.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: College of Life Science, Hebei Normal University, Shijiazhuang 050016, China. liujingze@mail.hebtu.edu.cn.
authors.forename: Jingze
authors.initials: J
authors.lastname: Liu
mesh_terms: D001173:Arthrobacter / Q000201:enzymology*; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology*; D002845:Chromatography; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D005942:Gluconates / Q000378:metabolism* / Q000494:pharmacology; D042843:Glucose 1-Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D012997:Solvents / Q000494:pharmacology; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature
chemical_list: D005942:Gluconates; D007477:Ions; D008670:Metals; D012997:Solvents; C008229:2-ketogluconate; D004492:Edetic Acid; D042843:Glucose 1-Dehydrogenase