Comparative determination of the rates of Kluyveromyces bulgaricus protoplast formation by the coulter counter technique and spectrophotometry

Abstract

AbstractKluyveromyces bulgaricus protoplast formation was studied by the determination of the remaining intact cells with the Coulter counter, the decrease of the absorbance of the cell suspension upon dilution into an hypoosmotic medium and the liberation of intracellular proteins after protoplast lysis. Due to the scattering of light by cells and ghosts, spectrophotometric readings were not enough specific and led to the underestimation of the reaction rates (about 40% less). The Coulter counter technique and the determination of the liberation of intracellular proteins gave the same rates over a large range of cell concentrations and in the presence of various cell lytic enzymes. The importance of corrected etermination of the time course of the reaction was illustrated by the good correlation obtained between protoplast formation and ATP turnover.