Evaluation of the Efficiency of Different Disruption Methods on Yeast Cell Wall Preparation for β-Glucan Isolation

Abstract

Selected methods for yeast cell disruption were evaluated to establish their suitability for cell wall preparation in the process of β-glucan isolation. The effect of different disruption methods on contents of total saccharides, β-glucans and proteins in the produced cell walls preparations was analyzed. The degree of cell wall purification from intracellular components was established on the basis of the ratio of solubilised material. The investigated methods included: cell exposure to hot water (autoclaving), thermally-induced autolysis, homogenization in a bead mill, sonication and their combinations. Experimental systems were prepared in water (pH 5.0 and pH 7.0) and Tris-HCl buffer (pH 8.0). The Saccharomyces cerevisiae yeast cell wall preparations with the highest degree of cytosol component release and purification of β-glucans were produced by 30 min of cell homogenization with zirconium-glass beads (0.5 mm in diameter). This was confirmed by the highest ratio of solubilised material (approx. 64%-67%). The thus-produced preparations contained ca. 60% of total saccharides, 13%-14% of β(1,3)/(1,6)-glucans, and approx. 35% of crude proteins. Similar results were obtained after autolysis coupled with bead milling as well as with sonication, but the time OPEN ACCESSMolecules 2014, 19 20942 required for these processes was more than 24 h. Homogenization in a bead mill could be valuable for general isolation procedures because allows one to eliminate the different autolytic activity of various yeast strains.

Characterization and emulsifying properties of extracts obtained by physical and enzymatic methods from an oenological yeast strain

“…Regardless of this, the yield of the physical method was higher than those previously obtained after heat treatment at 95 °C for 5 h . This suggests an important effect of the sonication applied, in agreement with results previously reported by Bzducha‐Wróbel et al . Furthermore, the high extraction yield obtained by the physical method could be due to the fact that, in order to simplify the protocols for future scale‐up applications, mannoproteins were not purified by ethanol precipitation as done by other authors .…”

“…A common feature visible in all the enzymatic extractions was the presence of small and spherical cell structures similar to those observed by Bzducha‐Wróbel et al . after extraction by physical methods . According to other authors, these spherical structures (Fig.…”

Evaluation of a novel method for measurement of intracellular calcium ion concentration in fission yeast

“…The cytolysis of fission yeast in pellet (D) was confi rmed in cells treated with HNO 3 at concentration of 36% or above. Various cell disruption strategies have been used to develop an efficient, low-cost, and effective method for release of intracellular products (Middelberg, 1995;Geciova et al, 2002;Ho et al, 2008;Klimek-Ochab et al, 2011;Bzducha-Wróbel et al, 2014). Unfortunately, the method of cell wall digestion reported in previous studies to might be too expensive.…”

δ15N from soil to wine in bulk samples and proline

“…The efficiency of the method in releasing intercellular yeast compound was evaluated by reading the absorbance value of the supernatant solution at wavelengths λ = 260 and 280 nm. The absorbance peaks at these wavelengths corresponded to absorption by nucleic acid and proteins respectively, these being the main intracellular constituents of S. cerevisiae …”