Enzyme Activity Measurement of N-Acylhexosamine Oxidase Using Spectrophotometric Assays

Creative Enzymes stays in the upmost class in the enzymology service industry, providing accurate and reliable activity assays for thousands of researchers globally. We have gained extensive experiences through years of enzyme activity testing. We are especially professional in method customization to create the most suitable assay based on the actual situations. Now, we brings all these advantages to the activity measurement for N-acylhexosamine oxidase.

N-acylhexosamine oxidase was first isolated from Pseudomonas sp. and has not been found in any other sources yet. This enzyme belongs to the family of oxidoreductases, in the subgroup which acts on the CH-OH group of donor with oxygen as the acceptor. N-acylhexosamine oxidase (EC 1.1.3.29) catalyzes the chemical reaction of N-acetyl-D-glucosamine, which is common and mainly in polysaccharides found in many natural products. The enzyme converts the sugar to N-acetyl-D-glucosaminate, meanwhile, the O₂ gas is reduced to hydrogen peroxide (H₂O₂). This enzyme also acts on other molecules with similar structure, including N-glycolylglucosamine, N-acetylgalactosamine and, more slowly, on N-acetylmannosamine. The systematic name of this enzyme class is N-acyl-D-hexosamine: oxygen 1-oxidoreductase, and it is also called N-acyl-D-hexosamine oxidase or N-acyl-beta-D-hexosamine:oxygen 1-oxidoreductase. Specially, the enzyme shows its activity throughout a wide pH range with the optimum pH at 8.0. The enzyme can be inhibited specifically by Zn²⁺. A preliminary study suggested that the enzyme could be used to measure the activity of N-acetyl-β-D-glucosaminidase (NAG), which is known for its application in the diagnosis of kidney injuries owning to diabetes or medication with antibiotics. There is no crystal structure of N-acylhexosamine oxidase, but studies showed that it is composed of four non-identical subunits. The lack of structural and biochemical information about N-acylhexosamine oxidase limits the detection and application of this enzyme. Clearly, correctly measuring its activity would be the first step to the path of product development. The enzymatic activity of N-acylhexosamine oxidase can be assayed through the detection of the O₂ consumption by oxygen electrode. However, Creative Enzymes displayed higher precision in the activity assay using a coupled enzyme system, in which the products can be easily quantified by spectrophotometry.

Figure: The reaction catalyzed by N-acylhexosamine oxidase.

Creative Enzymes provides unparalleled assay service for oxidoreductases. Our innovative and intelligent researchers always perform accurate activity tests. By combining our specialized research team and the advanced equipment, Creative Enzymes is able to supply the exact assay method to satisfy each client’s request.