title: Inhibitory kinetics of beta-N-acetyl-D-glucosaminidase from green crab (Scylla serrata) by zinc ion.
abstract: Heavy metal pollution such as chromium and zinc in the seawater has been increasing in recent years in the China Sea. Marine shellfish such as prawn and crab are sensitive to this pollution. Beta-N-acetyl-d-glucosaminidase (NAGase, EC 3.2.1.52) catalyzes the cleavage the oligomers of N-acetylglucosamine (NAG) into the monomer. In this study, taking p-nitrophenyl-N-acetyl-beta-d-glucosaminide (pNP-NAG) as substrate, the effects of Zn(2+) on NAGase from green crab ( Scylla serrata ) have been studied. The results showed that appropriate concentrations of zinc could lead to reversible inhibition on the enzyme, and the IC(50) has been estimated to be 0.5 +/- 0.012 mM. Furthermore, it has been shown that Zn(2+) could reduce the thermal stability of NAGase depending on the concentration of Zn(2+). The inhibitory kinetics of zinc on the enzyme in the appropriate concentrations has been studied using the kinetic method of substrate reaction. The inhibition model has been set up, and the rate constants have been determined. The results showed that Zn(2+) was a mixed-type inhibitor of NAGase and that it could combine at the free enzyme and the enzyme-substrate active sites.
authors.affiliation: Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystems, School of Life Sciences, Xiamen University, Xiamen 361005, China.
authors.forename: Ji-Ping
authors.initials: JP
authors.lastname: Zhang
authors.forename: Yong-Hua
authors.initials: YH
authors.lastname: Hu
authors.forename: Qin
authors.initials: Q
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Ye
authors.initials: Y
authors.lastname: Wang
authors.forename: Jiang-Hua
authors.initials: JH
authors.lastname: Yan
authors.forename: Qing-Xi
authors.initials: QX
authors.lastname: Chen
mesh_terms: D000118:Acetylglucosaminidase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D000818:Animals; D003386:Brachyura / Q000737:chemistry / Q000201:enzymology*; D020134:Catalytic Domain; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D007700:Kinetics; D015032:Zinc / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D000118:Acetylglucosaminidase; D015032:Zinc