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Optimized methodology for extraction of (1 → 3)(1 → 6)-β-d-glucan from Saccharomyces cerevisiae and in vitro evaluation of the cytotoxicity and genotoxicity of the corresponding carboxymethyl derivative
Authors:Marciane Magnani  Caroline M Calliari  Fernando C de Macedo Jr  Mateus P Mori  Ilce M de Syllos Clus  Raul JH Castro-Gomez
Institution:aDepartamento de Ciência e Tecnologia de Alimentos, Universidade Estadual de Londrina, Londrina, Paraná, Brazil;bDepartamento de Química, Universidade Estadual de Londrina, Londrina, Paraná, Brazil;cDepartamento de Biologia Geral, Universidade Estadual de Londrina, Londrina, Paraná, Brazil
Abstract:The cell wall of Saccharomyces cerevisiae is an important source of β-d-glucan, a glucose homopolymer with immunostimulant properties. The standard methodologies described for its extraction involve acid and alkaline washings, which degrade part of its glucose chains and reduce the final yield. In the present study, an optimized methodology for extraction of β-d-glucan from S. cerevisiae cells, involving sonication and enzyme treatment, with a yield of 11.08 ± 0.19%, was developed. The high-purity (1 → 3)(1 → 6)-β-d-glucan was derivatized to carboxymethyl-glucan (CM-G). In vitro tests with CM-G in Chinese hamster epithelial cells (CHO-k1) did not reveal any cytotoxic or genotoxic effects or influences of this molecule on cell viability. The method described here is a convenient alternative for the extraction of (1 → 3)(1 → 6)-β-d-glucan under mild conditions without the generation of wastes that could be potentially harmful to the environment.
Keywords:β  -Glucan  Saccharomyces cerevisiae  Sonication  Carboxymethyl-glucan (CM-G)  Comet assay
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