Structural stability of lipase from wheat germ in alkaline pH.

The present investigation shows the effect of alkaline pH on the structure-function relationship of lipase from wheat germ. There is a 70% decrease in lipase activity at pH 10.0, which decreases to 93% at pH 12.0 as compared to neutral pH activity (Rajendran et al. 1990). This change is shown to be as a result of loss of alpha-helical structure with a concomitant increase in aperiodic structure. The results with fluorescence spectra and tyrosyl ionization indicate gradual exposure of aromatic side chains of tyrosine and tryptophan to the bulk solvent along with the structural changes. The enzyme is in an extended form at alkaline pH with a volume change of - 1300 ml mol as also indicated by increase in reduced viscosity to 12.5 ml g and significant decrease in sedimentation coefficient. The kinetics of the reaction points to a cooperative pseudo first-order reaction as determined by stopped-flow kinetic analysis in the ultraviolet region. The inactivation mechanism appears to follow a two-step mechanism of a fast and a slow reaction.