Enzyme immobilization

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Spectrophotometric Determination of Enzymatically Generated Hydrogen Peroxide using Sol-Gel Immobilized Horseradish Peroxidase

Journal Title, Volume, Page: 
Talanta Volume 44, Issue 11, November 1997, Pages 2051–2058 In Honour of Professor George-Emil Baiulescu
Year of Publication: 
1997
Authors: 
Faida A. El-Essi
Chemistry Department, Faculty of Science, An-Najah National University, P.O. Box 707, Nablus Palestine
Ali Z.Abu Zuhri
Chemistry Department, Faculty of Science, An-Najah National University, P.O. Box 707, Nablus Palestine
Suleiman I. Al-Khalil
Chemistry Department, Faculty of Science, An-Najah National University, P.O. Box 707, Nablus Palestine
Current Affiliation: 
Department of Medical Laboratory Sciences, Faculty of Science, An-Najah National University, Palestine
Monzir S. Abdel-Latif
Chemistry Department, The Islamic University, P.O. Box 108, Gaza, Palestinian Authority
Preferred Abstract (Original): 

Peroxidase entrapment in different Sol-Gel matrices was successful. The enzyme did not show a decrease in activity for at least 2 months as well as storage at room temperature and dry condition for periods exceeding 3 weeks. It was evident that the enzymatic activity was a function in the type of the alkoxysilane precursor. In addition, the optimum temperature which resulted in maximum enzymatic activity was also dependent on the type of Sol-Gel matrix. Excellent results were obtained for the determination of glucose in serum samples using soluble glucose oxidase in conjunction with the Sol-Gel entrapped peroxidase. The enzymatically produced hydrogen peroxide is oxidized by the entrapped peroxidase yielding oxygen which oxidizes the faint blue variamine blue into the intensely violet colored species (the molar absorptivity is about 1.8 × 104 1 mol−1 cm−1). The characteristics of this chromogenic system as well as optimized conditions for its use in the spectrophotometric determination of enzymatically generated hydrogen peroxide were investigated. Excellent agreement between the results obtained by the proposed method and the widely used standard method, utilizing a commercial reagents kit, was always observed.

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