Solubilization of yeast cell-wall β-(1→3)-d-glucan by sodium hypochlorite oxidation and dimethyl sulfoxide extraction
Section snippets
. Introduction
Candida spp. is a medically important genus of fungi that induces disseminated candidiasis and candidemia in hospitalized, immunocompromised patients. The cell wall of Candida is mainly composed of two polysaccharides, mannan and β-glucan, and at least a part of the β-glucan, mainly the β-(1→3)-d-glucan moiety, is basically insoluble in H2O or NaOH and is quite difficult to extract [1], [2]. To analyse the architecture of the yeast cell-wall, traditional procedures, such as repeated acid and/or
Materials
All strains of Candida albicans, Candida parapsilosis, and Saccharomyces cerevisiae were purchased from the Institute for Fermentation, Osaka, maintained on Sabouraud agar (Difco, USA) at 25 °C and transferred once every 3 months. Sodium hypochlorite solution and sodium hydroxide were purchased from Wako Pure Chemical Industries, Ltd. The limulus G-test (Fungitec G test MK) and zymolyase (20T and 100T) were from Seikagaku Corp. (Tokyo), and distilled water (DIW) was from Otsuka Co., Ltd.
Preparation of cell-wall β-glucan by NaClO oxidation, followed by Me2SO extraction
Acetone-dried yeast cells were suspended in 0.1 M NaOH and oxidized with NaClO solution for 1 day at 4 °C. After the reaction was completed, the reaction mixture was centrifuged to collect a particulate substance, and the residue was washed extensively with water and dried with ethanol and acetone. During establishment of the preparation procedure, dialysis of the whole reaction mixture was performed instead of centrifugation. Components and yields were comparable in either method; thus we chose
. Discussion
We established a convenient, two-step, procedure to solubilize the yeast cell-wall β-(1→3)-d-glucan by combined use of NaClO oxidation and Me2SO extraction. This method was applied to several strains of Candida and Saccharomyces. The structures of all the products were essentially confirmed as β-(1→3)-d-glucan, but covalently bound with various amounts of the β-(1→6)-d-glucan moiety. The proportion of the β-(1→6)-d-glucan moiety was lower in examples submitted to harsher NaClO oxidation. The
Acknowledgements
The authors would like to express sincere thanks to Mr Y. Ohgoshi for excellent technical assistance.
References (26)
- et al.
Arch. Biochem. Biophys.
(1986) - et al.
Carbohydr. Res.
(1997) - et al.
Carbohydr. Res.
(1992) - et al.
FEMS Immunol. Med. Microb.
(1996) - et al.
Carbohydr. Res.
(1991) - et al.
Carbohydr. Res.
(1985) - et al.
FEMS Immunol. Med. Microb.
(1996) - et al.
Lancet
(1995) - et al.
Can. J. Chem.
(1967) - et al.
J. Gen. Microbiol.
(1976)
Microbiol. Immunol.
Biol. Pharm. Bull.
Biol. Pharm. Bull.
Cited by (130)
β-Glucan: A dual regulator of apoptosis and cell proliferation
2021, International Journal of Biological MacromoleculesFungal Polysaccharides
2021, Comprehensive Glycoscience: Second EditionDevelopment of a novel β-1,6-glucan–specific detection system using functionally-modified recombinant endo-β-1,6-glucanase
2020, Journal of Biological ChemistryCitation Excerpt :The β-1,6-glucan islandican from Penicillium islandicum (50), Agaricus brasiliensis–derived AgCAS (51) that is rich in β-1,6-glucan, and the branched β-1,3-glucan SCG from Sparassis crispa (52, 53) were prepared as described previously. Solubilized β-glucan purified from cell wall, CSBG from C. albicans NBRC 1385 (54), ASBG from A. niger NBRC 6342 (55), and the exopolysaccharide CAWS released from C. albicans NBRC 1385 (17) were prepared according to previous reports. Chitin oligomers were prepared through acetone precipitation after hydrolysis in concentrated hydrochloric acid as described previously (56).
Effect of yeast cell wall and (1→3)-β-D-glucan on transfer of aflatoxin from feed to milk in Saanen dairy goats
2019, Animal Feed Science and TechnologyCitation Excerpt :Briefly, yeast cells were treated by sequential steps of alcohol suspension, oxidation, extensive dialysis, oxidation, boiling, sonication, lyticase (SIGMA L4025, USA) exposure and freeze drying. Purity of βG was examined by the comparison of Curdlan (SIGMA C7821, USA), commercial form of pure βG, and βG hydrogen-1 nuclear magnetic resonance (1H NMR) spectroscopy signals (Ohno et al., 1999). For this purpose, Curdlan, as standard and βG was dissolved in Me2SO-d6, at 55 °C.
Production of low-molecular weight soluble yeast β-glucan by an acid degradation method
2018, International Journal of Biological Macromolecules