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

Molecules

Anna Bzducha-Wrl¹  Stanisᐪw Bᐪ៎jak¹  Anna Kawarska¹  Lidia Stasiak-Ró៊ńska¹  Iwona Gientka¹  Ewa Majewska² 

[1]Department of Biotechnology, Microbiology and Food Evaluation, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, Warsaw 02-776, Poland

[2] E-Mails:

[3]Department of Chemistry, Warsaw University of Life Sciences-SGGW, Nowoursynowska 159c, Warsaw 02-776, Poland, E-Mail:

关键词: β-glucan;    yeast cells wall;    bead mill;    autolysis;    sonication;    autoclaving;   

DOI  :  10.3390/molecules191220941

来源: mdpi

【 摘 要 】

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 (0.5mm 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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.