Physicochemical characteristics and anti-inflammatory activities of antrodan, a novel glycoprotein isolated from Antrodia cinnamomea mycelia.
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.
Research Institute of Biotechnology, Hungkuang University, 34 Chung-Chie Road, Shalu District, Taichung 433, Taiwan.
Chun-Hung
CH
Chiu
Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, 250 Wu-Hsing Street, Taipei 11031, Taiwan.
Chiung-Chi
CC
Peng
Department of Applied Food Technology, Hungkuang University, 34 Chung-Chie Road, Shalu District, Taichung 433, Taiwan.
Yaw-Bee
YB
Ker
Grape King Biotechnology Inc., Zhongli City, Taoyuan County 320, Taiwan.
Chin-Chu
CC
Chen
Research Institute of Biotechnology, Hungkuang University, 34 Chung-Chie Road, Shalu District, Taichung 433, Taiwan.
Arwen
A
Lee
Research Institute of Biotechnology, Hungkuang University, 34 Chung-Chie Road, Shalu District, Taichung 433, Taiwan.
Wan-Lin
WL
Chang
Research Institute of Biotechnology, Hungkuang University, 34 Chung-Chie Road, Shalu District, Taichung 433, Taiwan. ccchyau@sunrise.hk.edu.tw.
Charng-Cherng
CC
Chyau
Research Institute of Biotechnology, Hungkuang University, 34 Chung-Chie Road, Shalu District, Taichung 433, Taiwan. ypeng@seed.net.tw.
Robert Y
RY
Peng
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D055491:Antrodia / Q000737:chemistry*; D002241:Carbohydrates / Q000737:chemistry; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D005936:Glucans / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D009005:Monosaccharides / Q000737:chemistry; D025282:Mycelium / Q000737:chemistry*; D009569:Nitric Oxide / Q000096:biosynthesis; D014574:Uronic Acids / Q000737:chemistry
D000596:Amino Acids; D002241:Carbohydrates; D005656:Fungal Proteins; D005936:Glucans; D006023:Glycoproteins; D009005:Monosaccharides; D014574:Uronic Acids; D009569:Nitric Oxide