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Molecules 2014, 19(1), 22-40; doi:10.3390/molecules19010022
Article

Physicochemical Characteristics and Anti-Inflammatory Activities of Antrodan, a Novel Glycoprotein Isolated from Antrodia cinnamomea Mycelia

1,†, 2,†, 3, 4, 1, 1, 1,*  and 1,*
1 Research Institute of Biotechnology, Hungkuang University, 34 Chung-Chie Road, Shalu District, Taichung 433, Taiwan 2 Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, 250 Wu-Hsing Street, Taipei 11031, Taiwan 3 Department of Applied Food Technology, Hungkuang University, 34 Chung-Chie Road, Shalu District, Taichung 433, Taiwan 4 Grape King Biotechnology Inc., Zhongli City, Taoyuan County 320, Taiwan These authors contributed equally to this work.
* Authors to whom correspondence should be addressed.
Received: 11 October 2013 / Revised: 5 December 2013 / Accepted: 6 December 2013 / Published: 19 December 2013
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Abstract

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.
Keywords: Antrodia cinnamomea; mycelia; glycoprotein; antrodan; structure; anti-inflammatory activity Antrodia cinnamomea; mycelia; glycoprotein; antrodan; structure; anti-inflammatory activity
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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Chiu, C.-H.; Peng, C.-C.; Ker, Y.-B.; Chen, C.-C.; Lee, A.; Chang, W.-L.; Chyau, C.-C.; Peng, R.Y. Physicochemical Characteristics and Anti-Inflammatory Activities of Antrodan, a Novel Glycoprotein Isolated from Antrodia cinnamomea Mycelia. Molecules 2014, 19, 22-40.

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