title: Kinetics and mechanism of jack bean urease inhibition by Hg2+.
abstract: BACKGROUND
  Jack bean urease (EC 3.5.1.5) is a metalloenzyme, which catalyzes the hydrolysis of urea to produce ammonia and carbon dioxide. The heavy metal ions are common inhibitors to control the rate of the enzymatic urea hydrolysis, which take the Hg2+ as the representative. Hg2+ affects the enzyme activity causing loss of the biological function of the enzyme, which threatens the survival of many microorganism and plants. However, inhibitory kinetics of urease by the low concentration Hg2+ has not been explored fully. In this study, the inhibitory effect of the low concentration Hg2+ on jack bean urease was investigated in order to elucidate the mechanism of Hg2+ inhibition.
  
  
  RESULTS
  According to the kinetic parameters for the enzyme obtained from Lineweaver-Burk plot, it is shown that the Km is equal to 4.6±0.3 mM and Vm is equal to 29.8±1.7 μmol NH3/min mg. The results show that the inhibition of jack bean urease by Hg2+ at low concentration is a reversible reaction. Equilibrium constants have been determined for Hg2+ binding with the enzyme or the enzyme-substrate complexes (Ki =0.012 μM). The results show that the Hg2+ is a noncompetitive inhibitor. In addition, the kinetics of enzyme inhibition by the low concentration Hg2+ has been studied using the kinetic method of the substrate reaction. The results suggest that the enzyme first reversibly and quickly binds Hg2+ and then undergoes a slow reversible course to inactivation. Furthermore, the rate constant of the forward reactions (k+0) is much larger than the rate constant of the reverse reactions (k-0). By combining with the fact that the enzyme activity is almost completely lost at high concentration, the enzyme is completely inactivated when the Hg2+ concentration is high enough.
  
  
  CONCLUSIONS
  These results suggest that Hg2+ has great impacts on the urease activity and the established inhibition kinetics model is suitable.
authors.affiliation: Fuyang Normal College, College of Chemistry and Chemical Engineering, Fuyang, 236037, People's Republic of China. shenglq@fync.edu.cn.
authors.forename: Nana
authors.initials: N
authors.lastname: Du
authors.forename: Mingming
authors.initials: M
authors.lastname: Chen
authors.forename: Zhaodi
authors.initials: Z
authors.lastname: Liu
authors.forename: Liangquan
authors.initials: L
authors.lastname: Sheng
authors.forename: Huajie
authors.initials: H
authors.lastname: Xu
authors.forename: Shuisheng
authors.initials: S
authors.lastname: Chen