Research Institute of Biotechnology, Hungkuang University, 34 Chung-Chie Road, Shalu District, Taichung 433, Taiwan.
Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, 250 Wu-Hsing Street, Taipei 11031, Taiwan.
Molecules. 2013 Dec 19;19(1):22-40. doi: 10.3390/molecules19010022.
Antrodia cinnamomea (AC) is a unique fungus found inhabiting the rotten wood of Cinnamomum kanehirai. A submerged liquid culture of AC has been developed and its bioproducts have been used to meet the market demand for natural fruiting bodies. AC exhibits anti-inflammatory, antitumor, antioxidant, and immunomodulatory effects. Previously, we isolated polysaccharide AC-2 from AC mycelia by means of alkali extraction with subsequent acid precipitation and found it had a pronounced anti-inflammatory effect. In this study, a novel polysaccharide named "antrodan" was obtained by further purification of AC-2 using Sepharose CL-6B column chromatography. Antrodan exhibited a molecular weight of 442 kD and contained a particularly high content of uronic acid (152.6±0.8 mg/g). The protein content was 71.0%, apparently, higher than the carbohydrate content (14.1%), and thus antrodan was characterized as a glycoprotein. Its total glucan content was 15.65%, in which β-glucan (14.20%) was prominently higher than α-glucan (1.45%). Its FTIR confirmed the presence of β-linkages between sugars, and intramolecular amide bonds between sugars and amino acids. Its 1H-NMR spectrum showed that antrodan was a complex union of α- and β-glucans, which had (1→4)-linked α-Glcp and (1→3)-linked β-Glcp linkages to the carbohydrate chains via asparagine linked to protein site. Biologically, antrodan was confirmed to be totally non-detrimental to RAW 264.7 cell line even at dose as high as 400 μg/mL. It showed potent suppressing effect on the lipopolysaccharide-induced inflammatory responses in RAW 264.7 cell line. Moreover, antrodan significantly reduced the nitrogen oxide production at doses as low as 18.75 μg/mL.
樟芝（Antrodia cinnamomea，AC）是一种独特的真菌，寄生于肉桂树腐朽的木材中。现已开发出 AC 的液体深层发酵培养方法，并利用其生物产物来满足对天然子实体的市场需求。AC 具有抗炎、抗肿瘤、抗氧化和免疫调节作用。先前，我们通过碱提取和随后的酸沉淀从 AC 菌丝体中分离出多糖 AC-2，并发现其具有明显的抗炎作用。在这项研究中，我们通过 Sepharose CL-6B 柱层析进一步纯化 AC-2 得到了一种新型多糖，命名为“antrodan”。Antrodan 的分子量为 442 kD，含有特别高含量的糖醛酸（152.6±0.8 mg/g）。蛋白质含量为 71.0%，显然高于碳水化合物含量（14.1%），因此 antrodan 被定性为糖蛋白。其总葡聚糖含量为 15.65%，其中β-葡聚糖（14.20%）明显高于α-葡聚糖（1.45%）。其 FTIR 证实了糖之间存在β键，以及糖和氨基酸之间的分子内酰胺键。其 1H-NMR 图谱表明，antrodan 是α-和β-葡聚糖的复杂结合物，通过天冬酰胺与蛋白质结合位点连接，碳水化合物链上有（1→4）连接的α-Glcp 和（1→3）连接的β-Glcp 键。生物学研究表明，antrodan 即使在高达 400 μg/mL 的剂量下，对 RAW 264.7 细胞系也完全没有损害。它对 LPS 诱导的 RAW 264.7 细胞系炎症反应具有很强的抑制作用。此外，antrodan 在低至 18.75 μg/mL 的剂量下即可显著降低一氧化氮的产生。
