The mechanism for nonspecific lipase from rat pancreas

Purified nonspecific lipase from rat pancreas appears to mediate the followin reaction sequence: (2) HE +nB in equilibrium HEBn; (2) HEBn + S inequilibrium HEBnS; (3) HEBnS IN EQUILIBRIUM EBnS' + ROH; (4) EBnS' + h2o vector HEBn + R'COO- + H+, where E equils enzyme, B equils bile salt, and S equils R'COOR. Evidence is presented for the occurrence, sequence and reversibility or irreversibility of the above four reactions. Also discussed are the activation energies for both a "good" and a "poor" substrate, the correlation between V and subsitituent function (o-) for a series of substituted benzoic acid esters, and the effects of steric hindrance. Studies of the incorporation of label from H2(18)O further detail the hydrolytic mechanism, and the effects of the physical state in which substrate is presented to the enzyme (micelles, emulsion, crude bulk phase, etc.) are discussed.