This chapter describes a zymographic assay of oxidases which is based on a coupled peroxidase or hemin reaction. The enzymatic activity of oxidases (i.e., diamine oxidase/DAO, glucose oxidase, galactose oxidase) can be directly monitored on polyacrylamide gels containing horseradish peroxidase or hemin, in the presence of their specific substrates and ortho-phenylenediamine (OPDA), an oxidizable chromogen. In the presence of hydrogen peroxide, OPDA is oxidized to azo-aniline, which led to well-defined yellow-brown bands on gels, with intensities corresponding to the enzymatic activity of oxidases.
Mercury (Hg) has been recognized as a global pollutant with a toxic, mobile, and persistent nature. It adversely affects the ecosystem and human health. Already developed biosensors for Hg detection majorly suffer from poor sensitivity and specificity.
School of Chemistry and Chemical Engineering, Southeast University, Nanjing 211189, China; Center for Global Health, School of Public Health, Nanjing Medical University, Nanjing 211166, China; Jiangsu Province Engineering Research Center of Antibody Drug, Key Laboratory of Antibody Technique of National Health Commission, Nanjing Medical University, Nanjing 211166, China. Electronic address:
Due to the easy preparation, high stability and environmental friendliness, nanozymes are frequently used as promising substitutes to natural enzymes. However, the efficacy of nanozymes in biomedicine aspects is often hampered by their potential biotoxicity and limited bioavailability, which prompted structure adaption or carrier design to maximize nanozymes performance. Despite considerable efforts on carriers to deliver nanozymes efficiently, the systematic studies on enzyme-like activities of nanozymes related to platforms of nanozyme@carrier are sparse.
Iron-based nanozymes, recognized for their biocompatibility and peroxidase-like activities, hold promise as catalysts in tumor therapy. However, their concurrent catalase-like activity undermines therapeutic efficacy by converting hydrogen peroxide in tumor tissues into oxygen, thus diminishing hydroxyl radical production. Addressing this challenge, this study introduces the hemin-cysteine-Fe (HCFe) nanozyme, which exhibits exclusive peroxidase-like activity.
Nanozymes are nanomaterials with catalytic properties similar to enzymes and offer advantages over natural enzymes such as lower cost, ease of storage, and convenience for large-scale preparation. Thus, they have received increasing attention and are extensively applied in fields such as chemistry, sensing, food, environment, and medicine. Herein, a hemin-derived nanozyme (Hemin-CDs) was prepared using hemin as the precursor and used to substitute the natural horseradish peroxidase (HRP) for colorimetric detection.
Leptospirosis is a neglected zoonosis caused by a pathogenic spirochete Leptospira. Diagnosis of leptospirosis in the early stage is difficult and can be easily confused with other infections. The existing detection methods are considered chronophagous and labor-intensive.
