Concanavalin A photocross-linked affinity cryogels for the purification of horseradish peroxidase

Abstract

The present study describes an easy and efficient procedure for the purification of horseradish peroxidase from horseradish roots. For this purpose, supermacroporous cryogels having Concanavalin A were prepared by photosensitive cross-linking polymerization. Horseradish peroxidase binding and elution from the prepared cryogels were carried out changing various parameters such as initial peroxidase concentration and pH. The best binding performance was obtained at pH 7.0. The maximum horseradish peroxidase binding of the cryogels was found to be 3.85 mg g À1 cryogel. Horseradish peroxidase purification from crude extract resulted in 115.1-fold. SDS-PAGE analysis and circular dichroism measurements indicated that the horseradish peroxidase purification from horseradish roots was successfully carried out.

Enzymatically cross-linked peptide hydrogels for enhanced self-assembling capability and controlled drug release

“…Under the catalysis of HRP, the tyrosine residues in the Fmoc-GFYY peptide undergo a free radical cross-linking polymerization. [17][18][19] First, H 2 O 2 and HRP react to generate a high enzymatic state intermediate involving an Fe(IV) oxoferryl center and a porphyrinbased cation radical. The intermediate takes one hydrogen atom from the phenolic hydroxyl group of tyrosine to form a tyrosine free radical.…”

“…Under the action of HRP, the phenolic hydroxyl group of Tyr generates free radicals, which trigger the decomposition of hydrogen peroxide (H 2 O 2 ) and promote the formation of covalent bridges between aromatic rings, thereby forming further chemical cross-linking between Tyr-containing peptide molecules. [17][18][19] As a proof-of-concept, we designed a series of simple self-assembling peptides with various numbers of Tyr moieties (Scheme 1A): Fmoc-Gly-Phe-Tyr-Tyr (Fmoc-GFYY, bearing two Tyr), Fmoc-Gly-Phe-Phe-Tyr (Fmoc-GFFY, bearing one Tyr), and Fmoc-Gly-Phe-Phe-Gly (Fmoc-GFFG, bearing no Tyr as a control). As shown in Scheme 1B, the treatment of H 2 O 2 and HRP induces intermolecular chemical cross-linking of Fmoc-GFYY, which undergoes further fiber growth and cross-linking to form Fmoc-GFYY/enzymatically cross-linked hydrogels (Scheme 1B).…”

Development of a new affinity chromatography method for purification of horseradish peroxidase enzyme

“…Purification studies in the literature have been carried out using different chromatographic methods. 13,[15][16][17][41][42][43][44] Nevertheless, affinity chromatography is the most prominent chromatography technique. Upon comparing this technique to other chromatographic methods, the purification of enzymes in a single step and with a high yield is the main goal of many researchers.…”

In Situ-Forming Magnetic Fe3O4 Nanoroses on Defect-Controllable Mesoporous Graphene Oxide for Enzyme-Mimic Sensing

“…Glucose is an essential substrate as the metabolic component in the fermentation industry, food industry, and biomedical industry. , An electrochemical enzymatic sensor is one of the most applied methods for glucose detection because of its fast, accurate, and portable characteristics. − However, activity loss, storage limit, and high expense are always challenging for long-term analysis. − To address the above issue, recently, a nonenzymatic sensor is arousing increasing research interest due to its high performance and stability in usage. − As a core of nonenzymatic sensor, the electrode material enables mimicking of the redox function of flavin adenine dinucleotide of the enzyme to the detection target. , But current materials are still unable to absolutely substitute the enzyme on account of the work limits of a high alkaline environment, high catalytic potential, and low selectivity.…”