Inhibitory Effect of Dimethylsulfoxide on N-Acetyl-β-D-Glucosaminidase from Chinese Horseshoe Crab Tachypleus tridentatus
LIN Jiancheng, WU Jianhong, LIN Juanjuan
Key Laboratory of Ecological Environment and Information Atlas (Putian University), Fujian Provincial Key Laboratory of Ecology-Toxicological Effects & Control for Emerging Contaminants, College of Environmental and Biological Engineering, Putian University, Putian 351100, China
Abstract：The periodic molting of arthropods is closely related to N-acetyl-β-D-glucosaminidase (EC3.2.1.52, NAGase). NAGase was extracted from the viscera of Tachypleus tridentatus. The effect of dimethyl sulfoxide (DMSO) on NAGase was studied by using the kinetics method of enzymatic reaction. The results showed that DMSO had strong inhibitory effect on the NAGase, the enzyme activity decreased by 38.0% when its concentration reached 0.1 mol/L. The inhibitory effect of DMSO on NAGase was reversible. The kinetics showed that the inhibitory effect of DMSO solution on NAGase was the competitive and non-competitive mixed type. The inhibition constants of DMSO on free enzyme (KI) and on enzyme-substrate complex(KIS) were determined to be 0.200 mol/L and 1.063 mol/L, respectively. The value of KI was less thanthat of KIS, indicating a marked protective effect of the substrate on the inactivation of the enzyme. As the concentration of substrate increased, the inactivation of enzyme in DMSO turned to decrease. Meanwhile, UV absorption spectra and fluorescence emission spectra of NAGase were measured. The UV absorption of NAGase treated by DMSO was enhanced with increasing DMSO concentration, but the fluorescence emission peak intensity of the enzyme gradually decreased, their UV absorption peak and fluorescence emission peak of NAGase did not shift. The fluorescence emission of enzyme was almost quenched when the concentration of DMSO reached 0.65 mol/L. It showed that changes of enzyme conformation in the DMSO solvent led to inactivation of the NAGase. The above results confirmed that DMSO might further affect its molting by inhibiting the activity of NAGase from Tachypleus tridentatus.
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2023, Vol. 42 
