Inactivation Kinetics of beta-N-Acetyl-D-glucosaminidase from Green Crab (Scylla serrata) in Dioxane Solution
- 生命科学－已发表论文 
beta-N-Acetyl-D-glucosaminidase (NAGase, EC.220.127.116.11), which catalyzes the cleavage of N-acetylglucosamine polymers, is a composition of chitinase and cooperates with endochitinase and exo-chitinase to disintegrate chitin into N-acetylglucosamine (NAG). In this investigation, A NAGase from green crab (Scylla serrata) was purified and the effects of dioxane on the enzyme activity for the hydrolysis of p-Nitrophenyl-N-acetyl-beta-D-glucosaminide (pNP-NAG) were studied. The results show that appropriate concentrations of dioxane can lead to reversible inactivation of the enzyme and the inactivation is classified as mixed type. The value of IC(50), the dioxane (inactivator) concentration leading to 50% activity lost, is estimated to be 0.68%. The kinetics of inactivation of NAGase in the appropriate concentrations of dioxane solution has been studied using the kinetic method of the substrate reaction. The rate constants of inactivation have been determined. The results showed that k(+0) is much larger than k'(+0), indicating the free enzyme molecule is more fragile than the enzyme-substrate complex in the dioxane solution. It is suggested that the presence of the substrate offers marked protection of this enzyme against inactivation by dioxane.