The effect of calcium on protoplast release from species of Aspergillus

The effect of various osmotica and lytic enzyme treatments, on the release of stable protoplasts from Aspergillus niger and two strains of Aspergillus fumigatus, has been investigated. Most osmotica supported protoplast release at a concentration of 0.6 M except for CaCl2 and Ca(NO3)2. Supplementation of 0.6 M KCl with 0.2 M CaCl2 enhanced release from both species, but depended upon strain of organism, concentration of osmoticum and enzyme system employed. More protoplasts were released at low concentrations of KCl and/or when chitinase was present in the digestion medium. This enhancement effect was concerned with the Ca2+ ion since addition of Ca(NO3)2 also gave a higher yield and EDTA severely depressed protoplast release. Further investigations suggested that the effect of Ca2+ was not primarily acting as a supplementary osmoticum or concerned with protoplast membrane stabilization. Experiments using 45CaCl2 showed that the enhancement effect was probably due to Ca2+ affecting chitinase activity.