Co-Immobilization and Compartmentalization of Cholesterol Oxidase, Glucose Oxidase and Horseradish Peroxidase for Improved Thermal and H2o2 Stability

32 Pages Posted: 11 Jun 2022

See all articles by Manuel Pinelo

Manuel Pinelo

Technical University of Denmark

Katarzyna Jankowska

Technical University of Denmark

Sigyn Björk Sigurdardóttir

Technical University of Denmark

Jakub Zdarta

Poznan University of Technology

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Abstract

Glucose oxidase (GO X ) and cholesterol oxidase (CO X ) are enzymes with numerous practical applications, especially in biosensors for detecting glucose and cholesterol, respectively. One of the methods of determining glucose and cholesterol concentrations is colorimetry, where a cascade reaction of glucose and cholesterol oxidation by, respectively, GO X and CO X , is employed to produce H 2 O 2 followed by reaction of the chromogenic substrate 2,2’-azino-bis(3-ehylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) catalyzed by horseradish peroxidase (HRP) to give H 2 O as by-product. We proposed the application of electrospun fibers made of poly(D,L-lactide- co -glycolide) and commercial membrane UFX5 for co-immobilization of glucose oxidase or cholesterol oxidase with horseradish peroxidase combined with simultaneous co-immobilization by adsorption or by compartmentalization of enzyme separation using polyelectrolytes layers. The biosystems resulted in higher conversion efficiency of ABTS using immobilized enzymes at 45 °C and 65 °C compared to use of free enzymes. Moreover, after 5 catalytic cycles, biosystems based on UFX5 membrane and HRP or GO X or CO X immobilized by compartmentalization reached efficiencies of 89% and 34%, respectively, with less than 10% elution of initial amount of immobilized enzymes. In contrast, use of biosystems composed of electrospun fibers and co-immobilized enzymes resulted in enzyme elution of over 90% and lack of ABTS bioconversion.

Keywords: cholesterol oxidase, glucose oxidase, horseradish peroxidase, compartmentalization, co-immobilization

Suggested Citation

Pinelo, Manuel and Jankowska, Katarzyna and Sigurdardóttir, Sigyn Björk and Zdarta, Jakub, Co-Immobilization and Compartmentalization of Cholesterol Oxidase, Glucose Oxidase and Horseradish Peroxidase for Improved Thermal and H2o2 Stability. Available at SSRN: https://ssrn.com/abstract=4134284 or http://dx.doi.org/10.2139/ssrn.4134284

Manuel Pinelo (Contact Author)

Technical University of Denmark ( email )

Anker Engelunds Vej 1
Building 101A
Lyngby, 2800
Denmark

Katarzyna Jankowska

Technical University of Denmark ( email )

Anker Engelunds Vej 1
Building 101A
Lyngby, 2800
Denmark

Sigyn Björk Sigurdardóttir

Technical University of Denmark ( email )

Anker Engelunds Vej 1
Building 101A
Lyngby, 2800
Denmark

Jakub Zdarta

Poznan University of Technology ( email )

Pl. Marii Skłodowskiej-Curie 5
60-965
Poland

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