Immobilization of invertase via its carbohydrate moiety on macroporous glycidyl methacrylate

Abstract

Immobilization via carbohydrate moiety is suitable for immobilization of glycoenzymes because it has little effect on enzyme active site. Macroporous glycidyl methacrylate is better support for enzyme immobilization from the much more used polystyrene because of its lesser hydrophobicity. We found optimal conditions for invertase immobilization via its carbohydrate moiety on macroporous glycidyl methacryate by varying enzyme concentration. We obtained immobilized enzyme with specific activity of 5500 lU/g, which is the highest activity reported in the literature. Immobilized enzyme has Km=43 mmol/l, temperature optimum of 60ºC, and pH optimum between 3.5 and 5.5.

Imobilizacija preko šećernog dela je pogodna za glikoenzime zato što ima malo uticaja na aktivno mesto enzima. Makroporozni glicidil metakrilat je bolji nosač za imobilizaciju enzima od više korišćenog polistirena zbog svoje hidrofilnosti. Utvrđeni su optimalni uslovi za imobilizaciju invertaze preko šećernog dela na makroporozni glicidil metalkrilat variranjem koncentracije enizima. Dobijen je imobilizovani enzim specifične aktivnosti 5000 IU/g, što je najveća aktivnost do sada opisana u literaturi. Imobilizovani enzim je imao Km 43 mmol/l, temperaturni optimum na 60ºC i pH optimum između 4 i 5.