Purification and characterization of a novel glucooligosaccharide oxidase from Acremonium strictum T1.
A novel glucooligosaccharide oxidase was purified 495-fold from wheat bran culture of a soil-isolated Acremonium strictum strain T1 with an overall yield of 21%. This enzyme was composed of a single polypeptide chain with a molecular mass of 61 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and size-exclusion high-performance liquid chromatography. Its isoelectric point was pH 4.3-4.5. This enzyme contained 1 mol of FAD per mol of enzyme and showed absorption maxima at 274, 379 and 444 nm. This enzyme was stable in the pH range of 5.0 to 11.0 with an optimal reaction pH of 10.0. The optimal reaction temperature was 50 degrees C. It was stable up to 50 degrees C for 1 h at pH 7.8. This enzyme oxidized those oligosaccharides with glucose residue on the reducing end and each sugar residue jointed by alpha or beta-1,4 glucosidic bond. The relative activity of this enzyme toward maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose, maltoheptaose, lactose, cellobiose and glucose was 100:94:74:46:66:56:64:47:59. To our knowledge, this is the first report on the discovery of an glucooligosaccharide oxidase as judged from enzyme substrate specificity.
Lin
S F
SF
Institute of Biochemistry, National Yang-Ming Medical College, Taipei, Taiwan, R.O.C.
Yang
T Y
TY
Inukai
T
T
Yamasaki
M
M
Tsai
Y C
YC
D000164:Acremonium / Q000201:enzymology*; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D005656:Fungal Proteins / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008670:Metals / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013057:Spectrum Analysis; D013379:Substrate Specificity; D013696:Temperature
D005656:Fungal Proteins; D006023:Glycoproteins; D008670:Metals; D000429:Alcohol Oxidoreductases; C072250:glucooligosaccharide oxidase