The proportion of calcium-bound pectin in plant cell walls

The amount of pectin held in cell walls by ionic bonds only was determined by extraction with cyclohexanediamine tetraacetic acid (CDTA) at room temperature, to remove calcium ions without degrading the galacturonan chains. Enzymic degradation was avoided by extracting the cell walls with phenol-acetic acid-water during preparation. From cell walls of celery petioles, cress hypocotyls and tomato and cucumber pericarp CDTA extracted 64-100 mg g(-1) pectin, leaving 80-167 mg g(-1) uronic acid in the residue. An additional extraction at high ionic strength was used to make the galacturonan chains more flexible and thus detach any pectins held by steric interactions, but the amount released in this way was small. Most of the residual uronic acid polymers could be extracted by cold alkali and remained soluble on neutralisation, showing that it was not water-insolubility that prevented their extraction with CDTA. Covalent bonding was thought more likely.