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- Biophysical characterization of antibacterial compounds derived from pathogenic fungi Ganoderma boninense
- Syahriel Abdullah , Yoon Sin Oh , Min-Kyu Kwak , KhimPhin Chong
- J. Microbiol. 2021;59(2):164-174. Published online December 23, 2020
- DOI: https://doi.org/10.1007/s12275-021-0551-8
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Abstract
- There have been relatively few studies which support a link
between Ganoderma boninense, a phytopathogenic fungus
that is particularly cytotoxic and pathogenic to plant tissues
and roots, and antimicrobial compounds. We previously observed
that liquid-liquid extraction (LLE) using chloroformmethanol-
water at a ratio (1:1:1) was superior at detecting
antibacterial activities and significant quantities of antibacterial
compounds. Herein, we demonstrate that antibacterial
secondary metabolites are produced from G. boninense mycelia.
Antibacterial compounds were monitored in concurrent
biochemical and biophysical experiments. The combined
methods
included high performance thin-layer chromatography (HPTLC), gas chromatography-mass spectrometry (GC-MS), high-performance liquid chromatography (HPLC), fourier transform infrared (FTIR), and nuclear magnetic resonance (NMR) spectroscopy. The antibacterial compounds derived from mycelia with chloroform-methanol extraction through LLE were isolated via a gradient solvent elution system using HPTLC. The antibacterial activity of the isolated compounds was observed to be the most potent against Staphylococcus aureus ATCC 25923 and multidrug-resistant S. aureus NCTC 11939. GC-MS, HPLC, and FTIR analysis confirmed two antibacterial compounds, which were identified as 4,4,14α-trimethylcholestane (m/z = 414.75; lanostane, C30H54) and ergosta-5,7,22-trien-3β-ol (m/z = 396.65; ergosterol, C28H44O). With the aid of spectroscopic evaluations, ganoboninketal (m/z = 498.66, C30H42O6), which belongs to the 3,4-seco-27-norlanostane triterpene family, was additionally characterized by 2D-NMR analysis. Despite the lack of antibacterial potential exhibited by lanostane; both ergosterol and ganoboninketal displayed significant antibacterial activities against bacterial pathogens. Results provide evidence for the existence of bioactive compounds in the mycelia of the relatively unexplored phytopathogenic G. boninense, together with a robust method for estimating the corresponding potent antibacterial secondary metabolites.
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