title: STABILITY OF NATIVE AND MODIFIED α-GALACTOSIDASE OF Cladosporium cladosporioides.
abstract: By modifying carbohydrate component of glycoproteins it is possible to elucidate its role in manifestation of structural and functional properties of the enzyme. The comparison of activity and stability of the native and modified by oxidation with sodium periodate α-galactosidase of Cladosporium cladosporioides was carried out. To determine α-galactosidase activity the authors used n-nitrophenyl synthetic substrate, as well as melibiose; raffinose and stachyose. Modification of the carbohydrate component had a significant effect on catalytic properties of the enzyme. Both the reduction of V and enzyme affinity for natural and synthetic substrates were observed The native enzyme retained more than 50% ofthe maximum activity in the range of 20-60 °C, while for the modified enzyme under the same conditions that temperature range was 30-50 °C. The modified α-galactosidase demonstrated a higher thermal stability under neutral pH conditions. The residual activity of the modified α-galactosidase was about 30% when treated with 70% (v/v) methanol, ethanol and propanol. About 50% of initial activity was observed when 40% ethanol and propanol, and 50% methanol were used. It was shown that the modification of C. cladosporioides α-galactosidase by sodium periodate is accompanied by a significant decrease in enzyme activity and stability, probably caused by topological changes in the tertiary and quaternary structure of the protein molecule.
authors.forename: N V
authors.initials: NV
authors.lastname: Borzova
authors.forename: L D
authors.initials: LD
authors.lastname: Varbanets
mesh_terms: D002963:Cladosporium / Q000737:chemistry* / Q000201:enzymology; D057075:Enzyme Assays; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008553:Melibiose / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry; D016877:Oxidants / Q000737:chemistry; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000737:chemistry; D011487:Protein Conformation; D011887:Raffinose / Q000737:chemistry; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013696:Temperature; D000519:alpha-Galactosidase / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D005656:Fungal Proteins; D009844:Oligosaccharides; D016877:Oxidants; D010504:Periodic Acid; C005695:stachyose; D008553:Melibiose; C009288:metaperiodate; D000519:alpha-Galactosidase; D011887:Raffinose