title: 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 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.
authors.affiliation: Department of Biotechnology, Microbiology and Food Evaluation, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, Warsaw 02-776, Poland. anna_bzducha_wrobel@sggw.pl.
authors.forename: Anna
authors.initials: A
authors.lastname: Bzducha-Wróbel
authors.affiliation: Department of Biotechnology, Microbiology and Food Evaluation, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, Warsaw 02-776, Poland. stanislaw_blazejak@sggw.pl.
authors.forename: Stanisław
authors.initials: S
authors.lastname: Błażejak
authors.affiliation: Department of Biotechnology, Microbiology and Food Evaluation, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, Warsaw 02-776, Poland. kawarska.anna@gmail.com.
authors.forename: Anna
authors.initials: A
authors.lastname: Kawarska
authors.affiliation: Department of Biotechnology, Microbiology and Food Evaluation, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, Warsaw 02-776, Poland. lidia_stasiak_rozanska@sggw.pl.
authors.forename: Lidia
authors.initials: L
authors.lastname: Stasiak-Różańska
authors.affiliation: Department of Biotechnology, Microbiology and Food Evaluation, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, Warsaw 02-776, Poland. iwona_gientka@sggw.pl.
authors.forename: Iwona
authors.initials: I
authors.lastname: Gientka
authors.affiliation: Department of Chemistry, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, Warsaw 02-776, Poland. ewa_majewska@sggw.pl.
authors.forename: Ewa
authors.initials: E
authors.lastname: Majewska
mesh_terms: D002021:Buffers; D002473:Cell Wall / Q000737:chemistry*; D062610:Fungal Polysaccharides / Q000737:chemistry / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D013010:Sonication; D017550:Spectroscopy, Fourier Transform Infrared; D047071:beta-Glucans / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D002021:Buffers; D062610:Fungal Polysaccharides; D047071:beta-Glucans