Determining the kinetic and optimum characteristics of glucose oxidase immobilized on polyurethane
Turkish Journal of Biochemistry, cilt.49, sa.5, ss.621-628, 2024 (SCI-Expanded, Scopus, TRDizin)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 49 Sayı: 5
- Basım Tarihi: 2024
- Doi Numarası: 10.1515/tjb-2023-0214
- Dergi Adı: Turkish Journal of Biochemistry
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
- Sayfa Sayıları: ss.621-628
- Anahtar Kelimeler: glucose oxidase, immobilization, polyurethane, glucose, biosensor
- Sağlık Bilimleri Üniversitesi Adresli: Evet
Özet
Objectives: The primary aim of this study is to immobilize glucose oxidase (GOX) as a model enzyme using a simple and efficient method without the use of any toxic chemicals. This will enable the multiple reusability of GOX, thereby reducing the process cost. Furthermore, the objective is to determine the kinetic, optimum pH, and temperature parameters in comparison to soluble GOX. Methods: For immobilization, GOX at a concentration of 5 mg/mL was mixed with an equal volume of hypol prepolymer in a beaker. Immobilization was achieved by the formation of amide bonds between –NH2 groups on the enzyme surface and the prepolymer. The activities of polyurethane foam (PUF)-immobilized and soluble GOX were monitored using horse radish peroxidase enzyme as a second enzyme. 2,2′-Azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical ABTS+ generated as a result of the enzymatic reactions was detected at 420 nm. Results: The kinetic parameters for soluble GOX were Km=9.39 ± 1.13 mM and Vmax=9.14 ± 0.45 A.U sec−1 × 10−4, while for PUF-immobilized GOX Km=10.04 ± 1.30 mM and Vmax=9.09 ± 0.49 A.U sec−1 × 10−4. PUF-immobilized GOX displayed high performance during reuse experiments. PUF-immobilization led to the activity at higher temperatures and over a wide pH range compared to its soluble counterpart. Conclusions: GOX was successfully immobilized for the first time using PUF. This technique can be used in the healthcare and food sectors as well as in biosensor applications. In the future, when successfully applied to many enzymes of industrial value, it could offer significant advantages.