Laboratory for Immunopharmacology of Microbial Products School of Pharmacy Tokyo University of Pharmacy and Life Sciences 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan.
Laboratory for Immunopharmacology of Microbial Products, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan.
Int J Med Mushrooms. 2020;22(9):855-868. doi: 10.1615/IntJMedMushrooms.2020035888.
Mushroom is one of the major sources of β-glucan used in medical applications and traditional therapies. Thus, structure analysis and quantification of β-glucan content is crucial to evaluate medicinal mushrooms. Most studies concerning mushroom-derived β-glucan have been focused on β-1,3-glucans. However, recent investigations suggest that β-1,6 glucans have important roles for immunomodulating activity. Therefore, to elucidate the fine structure of various mushroom-derived β-glucans, we recently developed a novel β-1,6 glucan detection system using the function-modified recombinant β-1,6-glucanase. In this study, we performed an ELISA-like assay using modified β-1,6-glucanase and soluble dectin-1-Fc as the probes for β-1,6-glucan and β-1,3-glucan, respectively. Reactivity of ELISA to crude hot water extracts of edible mushrooms (Grifola frondosa, Agaricus bisporus, Pleurotus tuoliensis, P. eryngii, P. ostreatus, Hypsizygus marmoreus, and Lentinus edodes) was compared and L. edodes showed the strongest reactivity among them. An additional 19 different products of fresh L. edodes (shiitake mushroom) commercially available in Japan were also analyzed. This revealed limited differences in amounts of β-1,6-glucan and β-1,3-glucan in each shiitake mushroom. Furthermore, structural analysis of some purified β-glucans derived from medicinal mushrooms was performed, and their action for inducing tumor necrosis factor-α production from the murine bone marrow-derived dendritic cells was investigated. We found relation between reactivity to modified β-1,6-glucanase and its cytokine inducing activity. This assay could be useful for evaluating the strains of edible or medicinal mushrooms, which may be used as alternative medicines.
蘑菇是用于医学应用和传统疗法的β-葡聚糖的主要来源之一。因此，β-葡聚糖含量的结构分析和定量对于评估药用蘑菇至关重要。大多数关于蘑菇衍生β-葡聚糖的研究都集中在β-1,3-葡聚糖上。然而，最近的研究表明β-1,6-葡聚糖在免疫调节活性方面具有重要作用。因此，为了阐明各种蘑菇衍生β-葡聚糖的精细结构，我们最近开发了一种使用功能修饰的重组β-1,6-葡聚糖酶的新型β-1,6-葡聚糖检测系统。在这项研究中，我们使用修饰的β-1,6-葡聚糖酶和可溶性 dectin-1-Fc 作为β-1,6-葡聚糖和β-1,3-葡聚糖的探针，进行了 ELISA 样测定。比较了 ELISA 对食用蘑菇（灰树花、双孢蘑菇、金针菇、杏鲍菇、平菇、姬松茸和香菇）的热水粗提物的反应性，发现香菇的反应性最强。还分析了日本市售的 19 种不同的新鲜香菇（香菇）产品。这表明每个香菇中β-1,6-葡聚糖和β-1,3-葡聚糖的含量差异有限。此外，对一些来自药用蘑菇的纯化β-葡聚糖进行了结构分析，并研究了它们诱导鼠骨髓来源树突状细胞产生肿瘤坏死因子-α的作用。我们发现了对修饰的β-1,6-葡聚糖酶的反应性与其细胞因子诱导活性之间的关系。该测定法可用于评估可作为替代药物的食用或药用蘑菇的菌株。
