PEROXIDASE-LIKE ACTIVITY OF HEMOGLOBIN-BASED HYBRID MATERIALS AGAINST DIFFERENT SUBSTRATES AND THEIR ENHANCED APPLICATION FOR H2O2 DETECTION


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ALTINKAYNAK C., Turk M., Ekremoglu M., ÖZDEMİR N.

BULLETIN OF THE CHEMICAL SOCIETY OF ETHIOPIA, vol.35, no.3, pp.537-550, 2021 (Journal Indexed in SCI) identifier identifier

  • Publication Type: Article / Article
  • Volume: 35 Issue: 3
  • Publication Date: 2021
  • Doi Number: 10.4314/bcse.v35i3.6
  • Title of Journal : BULLETIN OF THE CHEMICAL SOCIETY OF ETHIOPIA
  • Page Numbers: pp.537-550
  • Keywords: Hydrogen peroxide, Hemoglobin, Hybrid Material, Colorimetric assay, HYDROGEN-PEROXIDE, NANOFLOWERS, HB, IMMOBILIZATION, PLATFORM

Abstract

Organic-inorganic hybrid nanoflowers method with unique properties are preferred than conventional immobilization methods for the past decade. Hereemoglobin-based hybrid material (FlbNFs@Cu) was synthesized under different experimental conditions (pH 5.0-9.0 and 0.01-0.50 mgmL(-1) of hemoglobin) obtaining a material size of 9-10 mu m. The encapsulation percentage and weight yield of HbNFs@Cu were determined as 100% and 6.7%, respectively. The peroxidase-like activities of the material against different substrates (ABTS and Guaiacol) were compared to free hemoglobin. The HLINFs@Cu hybrid structure exhibited V-max of 3.6995 EU/mg and a Michaelis-Menten constant (K-M) of 0.1357 mM/mL. The HbNFs@Cu hybrid material was then used to catalyze the oxidation of a peroxidase substrate ABTS to the pigmented product, which provided a colorimetric and spectrophotometric detection of H2O2. The linear operating range, detectable colorimetrically as H2O2 sensor, is 0.005-0.0042 mM, while the linear operating range, detectable spectrometically, is 0.003-0.0042 mM. The limits of detection of colorimetric and spectrophotometric sensors were 0.005 mM and 0.003 mM, respectively. Collectively, these results showed that HLINFs@Cu can be used as colorimetric biosensor for H2O2 in potential applications such as pharmaceutical food, biomedical, environmental, and industrial.