title: Hofmeister effects on activity and stability of alkaline phosphatase.
abstract: We have studied the effects on alkaline phosphatase of adding high concentrations (normally 1.0 M) of simple salts. It is necessary to allow for significant effects of salts on the extinction coefficient of the reaction product, and on the apparent pH of the buffer. Both activity and stability of the enzyme correlate well with the Hofmeister series in terms of the salt's kosmotropic/chaotropic properties, which are assessed by the Jones-Dole viscosity B coefficients (B(+) for cations and B(-) for anions). The catalytic activity or V(max)/K(m) of the enzyme showed a bell-shaped relationship with the (B(-)-B(+)) values of the salts present, being optimal with salts (such as NaCl, KCl, and KNO(3)) where the anion and cation have similar kosmotropic/chaotropic properties. This effect is believed to be enzyme-specific and relates to the impact of both cations and anions on the enzyme's surface pH, active site, and catalytic mechanism. Anions play a more predominant role than cations in affecting enzyme stability. The rate of irreversible thermal inactivation is strongly reduced by addition of kosmotropic anions like SO(4)(2-) (half-life increased from 8 to 580 min at 60 degrees C). This effect is general and the mechanism probably involves the ability of the ions to affect the water solvation layer around the enzyme molecule and to interact with both the surface and internal structure of the enzyme.
authors.affiliation: College of life sciences, Shenzhen University, Shenzhen 518060, Guangdong, China 518060. zyang@szu.edu.cn
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yang
authors.forename: Xiu-Ju
authors.initials: XJ
authors.lastname: Liu
authors.forename: Chao
authors.initials: C
authors.lastname: Chen
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Halling
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D002021:Buffers; D020134:Catalytic Domain; D002417:Cattle; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007422:Intestines / Q000201:enzymology; D007700:Kinetics; D012492:Salts
chemical_list: D002021:Buffers; D012492:Salts; D000469:Alkaline Phosphatase