Sakai Yumiko, Azuma Masayuki, Takada Yuki, Umeyama Takashi, Kaneko Aki, Fujita Tsuyoshi, Igarashi Koichi, Ooshima Hiroshi
Department of Applied Chemistry and Bioengineering, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan.
J Biosci Bioeng. 2007 Feb;103(2):161-6. doi: 10.1263/jbb.103.161.
The deletion of MCD4 leads to an increase in beta-1,6-glucan level and a decrease in glycosylphosphatidylinositol-anchored protein and mannan levels in the cell wall of Saccharomyces cerevisiae, suggesting that mcd4 deletion mutant (mcd4Delta) displays beta-glucans on the cell surface without a mannan cover. An observation of the cell surface of mcd4Delta cells and an examination of the effect of contact between mcd4Delta cells and mouse macrophages indicated that macrophages were activated by contact with mcd4Delta cells displaying beta-glucans on the cell surface. We further examined the effect of intraperitoneal ethanol-fixed mcd4Delta cells on the survival period of mice infected with Candida albicans. mcd4Delta cells prolonged the survival period, implying that mcd4Delta cells may enhance the immune function of mice via macrophage activation. Moreover, we examined the structures of beta-glucans (i.e., alkali- and acetic acid-insoluble beta-glucans) extracted from mcd4Delta with (13)C-NMR and the effect of extracted beta-glucans on TNF-alpha secretion from macrophages. The structures of the beta-glucans from mcd4Delta differed from those of wild type (WT); however, there was no difference in tumor necrosis factor-alpha (TNF-alpha) secretion level between beta-glucans from mcd4Delta and those from WT. The yield of purified beta-glucans obtained from dry cells of mcd4Delta was higher than that obtained from dry cells of WT. mcd4Delta may be a superior strain for the preparation of beta-glucans.
MCD4的缺失导致酿酒酵母细胞壁中β-1,6-葡聚糖水平升高,糖基磷脂酰肌醇锚定蛋白和甘露聚糖水平降低,这表明mcd4缺失突变体(mcd4Δ)在细胞表面展示β-葡聚糖而没有甘露聚糖覆盖。对mcd4Δ细胞的细胞表面观察以及对mcd4Δ细胞与小鼠巨噬细胞之间接触效果的检查表明,巨噬细胞通过与在细胞表面展示β-葡聚糖的mcd4Δ细胞接触而被激活。我们进一步研究了腹腔内乙醇固定的mcd4Δ细胞对白色念珠菌感染小鼠存活期的影响。mcd4Δ细胞延长了存活期,这意味着mcd4Δ细胞可能通过巨噬细胞激活增强小鼠的免疫功能。此外,我们用(13)C-NMR研究了从mcd4Δ中提取的β-葡聚糖(即碱和乙酸不溶性β-葡聚糖)的结构以及提取的β-葡聚糖对巨噬细胞分泌肿瘤坏死因子-α(TNF-α)的影响。mcd4Δ的β-葡聚糖结构与野生型(WT)不同;然而,mcd4Δ的β-葡聚糖与WT的β-葡聚糖之间在肿瘤坏死因子-α(TNF-α)分泌水平上没有差异。从mcd4Δ干细胞获得的纯化β-葡聚糖产量高于从WT干细胞获得的产量。mcd4Δ可能是制备β-葡聚糖的优良菌株。