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. 2020 Dec 23;59(2):164–174. doi: 10.1007/s12275-021-0551-8

Biophysical characterization of antibacterial compounds derived from pathogenic fungi Ganoderma boninense

Syahriel Abdullah 1,#, Yoon Sin Oh 2,#, Min-Kyu Kwak 2,✉, KhimPhin Chong 1,✉
PMCID: PMC7756191  PMID: 33355891

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.

Electronic Supplementary Material

Supplementary material is available for this article at 10.1007/s12275-021-0551-8 and is accessible for authorized users.

Keywords: antibacterial activity, disc diffusion assay, ergosterol, ganoboninketal, Ganoderma boninense, lanostane

Electronic Supplementary Material

12275_2021_551_MOESM1_ESM.pdf (511.8KB, pdf)

Supplementary material, approximately 511 KB.

Acknowledgments

This research was supported by the Ministry of Education of Malaysia (FRGS0384-SG-2/2014). Syahriel Abdullah and Yoon Sin Oh-Two contributed equally to this work and should be considered co-first authors.

Footnotes

Supplemental material for this article may be found at http://www.springerlink.com/content/120956.

Conflict of Interest

The authors declare that there are no competing interests associated with the manuscript.

These authors contributed equally to this work.

Contributor Information

Min-Kyu Kwak, Email: genie6@eulji.ac.kr.

KhimPhin Chong, Email: chongkp@ums.edu.my.

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12275_2021_551_MOESM1_ESM.pdf (511.8KB, pdf)

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