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P-Nitrophenyl penta-N-acetyl-beta-chitopentaoside as a Novel Synthetic Substrate for the Colorimetric Assay of Lysozyme

P-Nitrophenyl penta-N-acetyl-beta-chitopentaoside as a Novel Synthetic Substrate for the Colorimetric Assay of Lysozyme

F Nanjo, K Sakai, T Usui

J Biochem. 1988 Aug;104(2):255-8.

PMID: 2972699

Abstract:

p-Nitrophenyl beta-glycosides of N-acetylchitooligosaccharides (PNP-(GlcNAc)n n = 3-5) were examined as substrates for lysozyme [EC 3.2.1.17]. The enzyme released predominantly p-nitrophenyl N-acetyl-beta-D-glucosaminide (PNP-GlcNAc) from each substrate. Furthermore, the initial rate of PNP-GlcNAc formation in lysozyme-catalyzed hydrolysis of p-nitrophenyl penta-N-acetyl-beta-chitopentaoside (PNP-(GlcNAc)5) was about 350 and 25 times faster than those of p-nitrophenyl tri-N-acetyl-beta-chitotrioside (PNP-(GlcNAc)3) and p-nitrophenyl tetra-N-acetyl-beta-chitotetraoside (PNP-(GlcNAc)4), respectively. From these results, a new colorimetric assay method of lysozyme using PNP-(GlcNAc)5 as a substrate was developed on the basis of the determination of p-nitrophenol liberated from the substrate by lysozyme through a coupled reaction involving beta-N-acetylhexosaminidase (NAHase). The assay system gave a linear dose-response curve in the range of 2-120 micrograms of lysozyme in a 15-60 min incubation. The present assay was not significantly influenced by the ionic strength of the medium and was reproducible. This method using PNP-(GlcNAc)5 as a substrate was shown to be useful for lysozyme assay.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP59837148	p-Nitrophenyl galacto-N-bioside		59837-14-8	Price

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