Proteic Feeder Effect on Glucose Oxidase Aggregates Formation

Year 2008, Volume: 36 Issue: 4, 313 - 318, 01.12.2008

Yasemin İspirli Hakan Ayhan

Abstract

In this study, it is aimed to prepare cross-linked glucose oxidase GOD aggregates by using different proteic feeders.Bovine serum albumin BSA , gelatine and collagen were used separately as proteic feeders in the aggregate preparation process. The initial enzyme concentration was kept constant as 0.05 mg/ml. The amount of BSA varied from1 to 50 mg and the amount of gelatine and collagen were in the range of 5- 75 mg. A cross-linker, glutaraldehyde 2% v/v has been used in order to form GOD aggregates with proteic feeder. The highest immobilization efficiency wasfound when BSA feeder was used. Activities of both free and immobilised GOD were obtained by measuring theamount of hydrogen peroxide formed from glucose conversion, by using spectrophotometer. The maximum activitieswere obtained at following proteic feeder amounts: BSA 5 mg, collagen 60 mg, gelatine 50 mg. Kinetic parametersof native and immobilised enzyme have been calculated by using Lineweaver-Burk plots in the substrate range of0.01-1 M . The maximum reaction rates for free and immobilised enzyme with different proteic feeders, BSA, gelatine and collagen were calculated as 66.29, 12.22, 9.29, 4.53 mM/min-1, respectively. The corresponding saturationconstants of systems with the maximum activity were 7.9, 11.37, 22.58, 33.6 mM, respectively

Keywords

Cross-linked Glucose Oxidase aggregates, proteic feeder, bovine serum albumin, Gelatine, Collagen

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