LogP | -1.3 at 20℃ |
EPA chemical substance information | information provided by: ofmpeb.epa.gov (external link) |
Introduction | Horseradish Peroxidase (HRP, E.C.1.11.1.7) is a glycoprotein formed by the combination of enzyme protein and iron porphyrin. The enzyme is composed of multiple isozymes with a molecular weight of 40kDa and an isoelectric point of pH 9.0. The maximum absorption spectra of HRP prosthetic group and enzyme protein are 403nm and 275nm respectively, so people usually use RZ value (I. E., the ratio of OD403nm /OD275nm) to express the purity of the enzyme, the higher the purity. |
action | lactoperoxidase is present in the mammary gland, saliva and tear glands of mammals and their respective secretions, namely milk, saliva and tears. It is one of the common constituents of bovine and human milk and is present in all mammalian milks that have been tested to date. Lactoperoxidase is the first secreted peroxidase found, and plays an important role in protecting the mammary glands of lactating women and the intestinal tract of newborns from pathogenic microorganisms. It is part of the body's defense system. The chemical and immunological properties of peroxidase in the mammary gland, saliva and lacrimal gland are similar. |
Use | 1. Peroxidase (horseradish) is one of the most widely used labeling enzymes in ELISA, mainly because of its easy preparation, relatively low price, stable nature, heat resistance and organic solvents, there is little loss of activity upon coupling to the antigen or antibody. It is also commonly used with sewage treatment. Industrial wastewater contains a large number of phenol, bisphenol A and other classification and aromatic amine compounds, and other environmental pollution, it can be these pollutants as a substrate, oxidation into free radicals, free radicals in the aggregation of integrated precipitation polymer, thus greatly reducing the pollution to the environment. It has been studied in recent years as a food additive. Because of its non-toxic and harmless, reaction conditions, temperature, reaction is easy to control, in recent years has been studied for food preservation, detoxification and detection and other fields. 4. Detection field: it can be used in combination with glucose oxidase to detect glucose content. used in biochemical research, it is a major enzyme used in enzyme labeling. It is used in medicine to determine the content of glucose and galactose in biological fluids, and is used as a diagnostic enzyme. |
synthesis method | (1) horseradish peroxidase crude extract was obtained by crushing, leaching and centrifugation of horseradish peroxidase;(2) the crude extract of horseradish peroxidase was separated by ultrafiltration using an ultrafiltration membrane with a molecular weight cut-off of 70kDa-100kDa to obtain horseradish peroxidase ultrafiltrate;(3) the horseradish peroxidase ultrafiltration solution is ultrafiltered and concentrated with an ultrafiltration membrane with a molecular weight cut-off of 1-30kDa to obtain a horseradish peroxidase concentrate;(4) the horseradish peroxidase concentrate is dehydrated to obtain horseradish peroxidase. |
detection method | a rapid and quantitative detection method for peroxidase in pork, the detection method comprises the following steps:(1) preparing a series of standard enzyme solutions with horseradish peroxidase as the standard enzyme;(2) using carbamide peroxide as the reaction substrate, 5 '-Tetramethylbenzidine as chromogenic reagent, with sodium dihydrogen phosphate buffer as buffer solution, respectively, the standard enzyme solution prepared in step (1), the test reaction temperature is 20-30 ℃, the reaction pH value is 4.0-5.0, and the reaction time is 30-90S. After the reaction, dilute sulfuric acid solution is added to terminate the reaction to obtain the reaction product solution;(3) the reaction product solution was placed in the ultraviolet spectrophotometer, and the OD value was measured at the absorption wavelength of 450nm, and the standard curve of horseradish peroxidase concentration-OD value was obtained. The OD value of the reaction product solution after the peroxidase reaction in the pork to be tested was tested, and the concentration of peroxidase in the pork was calculated according to the standard curve. |
production method | with horseradish as raw material, after water extraction, it is fractionated by ammonium sulfate for the first time, and purified by calcium phosphate gel, then the product was refined by ethanol classification, ammonium sulfate second classification and crystallization. |