Purification and characterization of the glucoside 3-dehydrogenase produced by a newly isolated Stenotrophomonas maltrophilia CCTCC M 204024.
A soluble glucoside 3-dehydrogenase (G3DH) from Stenotrophomonas maltrophilia CCTCC M 204024, recently isolated from wheat soil in our laboratory, was purified to 37.4-fold with a yield of 24.7% and was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a molecular mass of 66 kDa. 2,6-Dichlorophenolindophenol (DCPIP) and ferricyanide were able to act as artificial electron acceptors for the enzyme. The optimal pH of G3DH was in the range of 6.0-7.0 in the presence of DCPIP. The enzyme was stable in the pH range of 4.4-10.6 and was sensitive to heat. G3DH exhibited extremely broad substrate specificity by converting many sugars to their corresponding 3-ketoglucosides. They produced a characteristic spectrum by alkaline treatment with a peak at 340 nm. The apparent Km values for validoxylamine A and D: -glucose were 8.3 and 1.1 mM, respectively. Cu2+, Ag2+, and Hg2Cl2 inhibited the activity of G3DH.
Zhang
Jian-Fen
JF
Institute of Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
Zheng
Yu-Guo
YG
Xue
Ya-Ping
YP
Shen
Yin-Chu
YC
D005948:Glucose Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D012988:Soil Microbiology; D020615:Stenotrophomonas maltophilia / Q000201:enzymology* / Q000254:growth & development / Q000302:isolation & purification*; D013379:Substrate Specificity; D013696:Temperature
D005948:Glucose Dehydrogenases; C051543:glucoside 3-dehydrogenase