A kinetic study of the hydrolysis of the diester dimethyl cis-cyclohex-4-en
e-1,2-dicarboxylate, to the (1S,2R)-monoester, catalysed by the enzyme Pig
Liver Esterase (PLE) was performed. The effects of the most relevant parame
ters that influence the enzymatic conversion were studied, such as pH, temp
erature and concentration of substrate and reaction products. It was conclu
ded that the pH at which the enzyme exhibits a maximum activity is pH 7. At
25 degrees C PLE presents a better long-term stability and enantioselectiv
ity than at higher temperatures, although the reaction rate is slower. The
kinetic results obtained are well described by the Michaelis-Menten equatio
n, although a slight deviation to this model was observed for low substrate
concentrations. Methanol, a co-product of the enzymatic hydrolysis, was fo
und to act as a non-competitive inhibitor of the reaction. The Michaelis-Me
nten parameters were determined and a comprehensive kinetic model, which al
ready accounts for methanol inhibition, is presented. (C) 2000 Society of C
hemical Industry.