2-Keto-D-Gluconate-Yielding Membrane-Bound D-Glucose Dehydrogenase from Arthrobacter globiformis C224: Purification and Characterization

• Main Authors: Qing Xue, Zhuan Wei, Wenjing Sun, Fengjie Cui, Silian Yu, Qiang Zhou, Jingze Liu
• Format: Article
• Language: English
• Published: MDPI AG 2015-01-01
• Series: Molecules
• Subjects: Arthrobacter globiformis; 2-ketogluconic acid (2KGlcA); D-glucose dehydrogenase (GlcDH); purification; enzymatic properties
• Online Access: http://www.mdpi.com/1420-3049/20/1/846

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.