Beta-galactosidase monitoring by a biosensor based on Clark electrode: its optimization, characterization and application.
beta-Galactosidase is an hydrolase enzyme that catalyzes the hydrolysis of beta-galactosides into monosaccharides. Substrates of different beta-galactosidases include ganglioside GM1, lactosylceramides, lactose, and various glycoproteins. A novel aspect of the activity determination of beta-galactosidase was presented. A glucose oxidase biosensor based on Clark electrode was utilized in order to monitor beta-galactosidase. Immobilization of glucose oxidase was made by gelatin and glutaraldehyde as cross-linker. Several parameters such as glucose oxidase activity, gelatin amount, and glutaraldehyde percentage for cross-linking were optimized. The most important parameter, lactose concentration in working buffer was studied in detail. Optimum temperature, thermal stability, optimum pH, buffer system and its concentration effect on the biosensor system, repeatability, reproducibility, and storage and operational stabilities of the biosensor were identified. A linear detection range for beta-galactosidase was observed between 9.4 x 10(-5) and 3.2 x 10(-2)U/ml. Finally, beta-galactosidase activity in artificial intestinal juice was investigated by the biosensor and the results obtained were compared with a reference spectrophotometric method.
Sezgintürk
Mustafa Kemal
MK
Ege University, Faculty of Science, Biochemistry Department, 35100 Bornova-Izmir, Turkiye. msezginturk@hotmail.com
Dinçkaya
Erhan
E
D015374:Biosensing Techniques / Q000295:instrumentation*; D004563:Electrochemistry / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005949:Glucose Oxidase / Q000737:chemistry*; D008839:Microelectrodes; D001616:beta-Galactosidase / Q000032:analysis*
D005949:Glucose Oxidase; D001616:beta-Galactosidase