Identification and Isolation of an Intermediate Metabolite with Dual Antioxidant and Anti-Proliferative Activity Present in the Fungus Antrodia cinnamomea Cultured on an Alternative Medium with Cinnamomum kanehirai Leaf Extract

Wen-Wen Zeng; Tsan-Chi Chen; Cheng-Huan Liu; Sheng-Yang Wang; Jei-Fu Shaw; Yu-Ting Chen

doi:10.3390/plants10040737

Identification and Isolation of an Intermediate Metabolite with Dual Antioxidant and Anti-Proliferative Activity Present in the Fungus Antrodia cinnamomea Cultured on an Alternative Medium with Cinnamomum kanehirai Leaf Extract

Plants (Basel). 2021 Apr 9;10(4):737. doi: 10.3390/plants10040737.

Authors

Wen-Wen Zeng¹, Tsan-Chi Chen², Cheng-Huan Liu^{1

3}, Sheng-Yang Wang⁴, Jei-Fu Shaw⁵, Yu-Ting Chen^{1

6}

Affiliations

¹ Institute of Genomics and Bioinformatics, National Chung Hsing University, Taichung 40227, Taiwan.
² Department of Ophthalmology, Far Eastern Memorial Hospital, Banqiao District, New Taipei City 22060, Taiwan.
³ Department of Food Science and Biotechnology, National Chung Hsing University, Taichung 40227, Taiwan.
⁴ Department of Forestry, National Chung Hsing University, Taichung 40227, Taiwan.
⁵ Department of Biological Science & Technology, I-Shou University, Kaohsiung 84001, Taiwan.
⁶ Ph.D. Program in Medical Biotechnology, National Chung Hsing University, Taichung 40227, Taiwan.

Abstract

The fungus Antrodia cinnamomea has been used as a folk medicine for various diseases, especially cancer. When A. cinnamomea is cultured on the original host, an endangered woody plant Cinnamomum kanehirai Hayata, the fungus produces more active ingredients, but its growth is slow. Here, C. kanehirai leaf ethanol extract (KLEE) was used as a substitute for C. kanehirai wood to culture A. cinnamomea on solid medium to shorten the culture period and produce active metabolites en masse. The antioxidant activities of methanol extracts from A. cinnamomea cultured on KLEE (MEAC-KLEE) were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging effect, reducing power, and ferrous ion-chelating effect, and the effective concentration (EC₅₀) values were 0.27, 0.74, and 0.37 mg mL^-1, respectively. MEAC-KLEE exhibited specific anti-proliferative activity against a non-small-cell lung cancer cell line (A549) by Annexin V assay. A secondary metabolite (2,4-dimethoxy-6-methylbenzene-1,3-diol, DMMB) present in the extract (MEAC-KLEE) was purified by high-performance liquid chromatography (HPLC) and identified by nuclear magnetic resonance (NMR) spectra. DMMB exhibited moderate antioxidant activity against DPPH radicals and reducing power, with EC₅₀ values of 12.97 and 25.59 μg mL^-1, respectively, and also induced apoptosis in A549 cells. Our results provide valuable insight into the development of DMMB for nutraceutical biotechnology.

Keywords: 2,4-dimethoxy-6-methylbenzene-1,3-diol; Antrodia cinnamomea; Cinnamomum kanehirai; anti-proliferative; antioxidant.

Abstract

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