KINETIC COMPARISONS OF TRIMER-GENERATING PECTATE AND ALGINATE LYASES BY REVERSED-PHASE ION-PAIR LIQUID-CHROMATOGRAPHY

Reversed-phase ion-pair liquid chromatography has been applied to the kinetic analysis of trimer-generating pectate lyases secreted by Clostridium populeti, Lachnospira multiparus, and two different strains of Erwinia chrysanthemi. A trimer-generating alginate lyase secreted by a marine bacterium was also analyzed and compared. A common feature of these enzymes is the initial formation of trimer, tetramer, pentamer, and larger oligouronates. Dimer can be formed from the later eliminative cleavage of pentamer and larger oligouronates. A general depolymerizing process involving an endolytic, followed by an exolytic mechanism, has been ascribed to these enzymes, with subtle differences noted with respect to the quantities of products formed with time.