title: Purification and characterization of an extremely stable glucose isomerase from Geobacillus thermodenitrificans TH2.
abstract: The D-glucose/D-xylose isomerase was purified from a thermophilic bacterium, Geobacillus thermodenitrificans TH2, by precipitating with heat shock and using Q-Sepharose ion exchange column chromatography, and then characterized. The purified enzyme had a single band having molecular weight of 49 kDa on SDS-PAGE. In the presence of D-glucose as a substrate, the optimum temperature and pH of the enzyme were found to be 80 degrees C and 7.5, respectively. The purified xylose isomerase of G. thermodenitrificans TH2 was extremely stable at pH 7.5 after 96 h incubation at 4 degrees C and 50 degrees C. When the thermal stability profile was analyzed, it was determined that the purified enzyme was extremely stable during incubation periods of 4 months and 4 days at 4 degrees C and 50 degrees C, respectively. The K(m) and V(max) values of the purified xylose isomerase from G. thermodenitrificans TH2 were calculated as 32 mM and 4.68 micromol/min per mg of protein, respectively. Additionally, it was detected that some metal ions affected the enzyme activity at different ratios. The enzyme was active and stable at high temperatures and nearly neutral pHs which are desirable for the usage in the food and ethanol industry.
authors.forename: L
authors.initials: L
authors.lastname: Konak
authors.forename: Y
authors.initials: Y
authors.lastname: Kolcuoğlu
authors.forename: E
authors.initials: E
authors.lastname: Ozbek
authors.forename: A
authors.initials: A
authors.lastname: Colak
authors.forename: B
authors.initials: B
authors.lastname: Ergenoglu
mesh_terms: D019747:Aldose-Ketose Isomerases / Q000737:chemistry / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange; D004795:Enzyme Stability; D056506:Geobacillus / Q000737:chemistry* / Q000201:enzymology; D005947:Glucose / Q000737:chemistry; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D014994:Xylose / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D014994:Xylose; D019747:Aldose-Ketose Isomerases; C023298:xylose isomerase; D005947:Glucose