Immobilized and Free Apoplastic Pectinmethylesterases in Mung Bean Hypocotyl

Abstract

The nature and the action pattern of apoplastic pectinmethylesterase (PME) isoforms were investigated in mung bean [Vigna radiata (L.) Wilzeck] hypocotyls. Successive extractions of neutral and alkaline PME isoforms present in hypocotyl native cell walls (referred to as PE1, PE2, PE3, PE4, with increasingly basic isoelectric points) revealed that solubilization of PE1, PE2, and PE4 did not induce any significant decrease in the cell-wall-bound PME activity. The in vitro de-esterification occurring when isolated cell walls were incubated with pectin resulted, then, from the activity of PE3. In addition, pH control of PME activity was shown to be much stronger for enzymes bound to cell walls, in their native state or reintroduced after solubilization, than for enzymes in solution. Mature cell walls showed much more activity than young cell walls, and were relatively enriched in two acidic PME isoforms missing in young cell walls. One acidic PME was also detected in the extracellular fluid. The acidic and neutral isoforms that could be easily transferred from their binding sites to their substrate might be those involved in the demethylation process developing along the mung bean hypocotyl.