A novel NAD-dependent dehydrogenase, highly specific for 1,5-anhydro-D-glucitol, from Trichoderma longibrachiatum strain 11-3.
A novel NAD-dependent dehydrogenase highly specific for 1,5-anhydro-D-glucitol (1,5-AG) was found in the cell extract of an imperfect fungus, Trichoderma longibrachiatum strain 11-3. This fungus used 1,5-AG as a sole carbon source for growth and transformed 1,5-AG into glucose. 1,5-AG dehydrogenase (AGH) was purified to homogeneity, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular mass of the purified enzyme was estimated to be 36 and 141 kDa by SDS-PAGE and by gel filtration, respectively, suggesting that the enzyme was homotetrameric. The enzyme was highly specific for 1,5-AG and did not exhibit activity with any sugar or sugar alcohol tested in this study other than 1,5-AG. A linear relationship between the initial rate of the enzyme reaction and the concentration of 1,5-AG at the physiological level was observed. The presence of glucose in abundance did not interfere with the relationship. The optimum temperature for the enzyme reaction was 50 degrees C, and the enzyme was stable at temperatures up to 70 degrees C. These results suggested that AGH is a novel enzyme and is useful for specifically diagnosing diabetes mellitus.
Yoshida
Nobuyuki
N
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara 630-0192, Japan. yoshidan@bs.aist-nara.ac.jp
Uchida
Etsuko
E
Katsuragi
Tohoru
T
Tani
Yoshiki
Y
D003847:Deoxyglucose / Q000097:blood / Q000378:metabolism*; D003920:Diabetes Mellitus / Q000097:blood / Q000175:diagnosis; D004795:Enzyme Stability; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D012988:Soil Microbiology; D013379:Substrate Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D013696:Temperature; D014242:Trichoderma / Q000201:enzymology* / Q000302:isolation & purification
C006584:1,5-anhydroglucitol; D003847:Deoxyglucose; C476095:1,5-anhydro-D-glucitol dehydrogenase; D013401:Sugar Alcohol Dehydrogenases