The Influence of Isoenzyme Composition and Chemical Modification on Horseradish Peroxidase@ZIF-8 Biocomposite Performance

Abstract

Many articles in the literature deal with horseradish peroxidase (HRP) biomineralization, but none pay attention to the isoenzyme composition of commercial HRP or the influence of the carbohydrate component of the protein molecule on the biomineralization process. To study the impact of these factors, we performed periodate oxidation of commercial HRP and a purified HRP-C isoform for biomineralization within ZIF-8. With purified HRP, enzyme@ZIF-8 biocomposites with higher activity were obtained, while periodate oxidation of the carbohydrate component of both commercial HRP and purified HRP-C yields biocomposites with very high activity in acetate buffer that does not degrade the ZIF-8 structure. Using acetate instead of phosphate buffer can prevent the false high activity of HRP@ZIF-8 biocomposites caused by the degradation of ZIF-8 coating. At the same time, purification and especially oxidation of the carbohydrate component of enzymes prior to biomineralization lead to significantly improved activity of the biocomposites.

Support Materials of Organic and Inorganic Origin as Platforms for Horseradish Peroxidase Immobilization: Comparison Study for High Stability and Activity Recovery

“…It is possible to observe distinct clusters of enzymes on the surface of the support after immobilization (Figure 3b), which is confirmed by the increased average particle size of the resulting biocatalytic system (approximately 440 nm). ZIF-8 is an attractive material for immobilizing enzymes, as demonstrated in a study by Stanišić et al [40], which showed the successful immobilization of HRP on this support mainly due to well-fitted particles. Figure 3c presents the morphology of pure MWCNTs.…”

State of the Art Technologies for High Yield Heterologous Expression and Production of Oxidoreductase Enzymes: Glucose Oxidase, Cellobiose Dehydrogenase, Horseradish Peroxidase, and Laccases in Yeasts P. pastoris and S. cerevisiae

“…Horseradish peroxidase (EC 1.11.1.7) belongs to the group of OXR. This glycoprotein, with 44 kDa, contains at least 15 distinct isoenzyme forms, the most common and, thus, most studied of which is isoenzyme C1A [15]. It was determined that the tertiary structure of HRP includes two domains formed by ten α-helices, four disulfide bonds, two Ca 2+ ions per molecule, and the prosthetic heme group [16][17][18].…”

In Situ Immobilization of Multi-Enzymes for Enhanced Substrate Channeling of Enzyme Cascade Reactions: A Nanoarchitectonics Approach by Directed Metal–Organic Frameworks

“…3.5 Å) due to their high surface area, good stability, and room temperature synthesis conditions in the water phase. , However, when enzymes such as horseradish peroxidase (HRP), glucose oxidase (GOx), catalase, and lipase were immobilized within ZIF-8 structures, they demonstrated low activity (<50%) in comparison to free enzymes. Recent studies showed that growing MOFs can be used for in situ immobilization of enzymes via approaches such as co-precipitation and biomineralization. − These approaches allow rapid formation of enzyme@MOF composites by a one-step reaction with high stability and activity. ,, Furthermore, a peptide-modulated MOF (ZIF-8) was used to immobilize HRP in situ via electrostatic interaction, with enhanced activity . However, beyond the MOF-based host matrix materials, the understanding of a confined enzyme environment and the mechanism of conformation and orientation changes are urgently needed …”