- Biophysical characterization of antibacterial compounds derived from pathogenic fungi Ganoderma boninense
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Syahriel Abdullah , Yoon Sin Oh , Min-Kyu Kwak , KhimPhin Chong
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J. Microbiol. 2021;59(2):164-174. Published online December 23, 2020
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DOI: https://doi.org/10.1007/s12275-021-0551-8
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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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Citations
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- Ganoderma boninense mycelia for phytochemicals and secondary metabolites with antibacterial activity
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Syahriel Abdullah , Se-Eun Jang , Min-Kyu Kwak , KhimPhin Chong
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J. Microbiol. 2020;58(12):1054-1064. Published online December 2, 2020
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DOI: https://doi.org/10.1007/s12275-020-0208-z
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1,065
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Abstract
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Antiplasmodial nortriterpenes with 3,4-seco-27-norlanostane
skeletons, almost entirely obtained from fruiting bodies, represent
the main evidential source for bioactive secondary
metabolites derived from a relatively unexplored phytopathogenic
fungus, Ganoderma boninense. Currently lacking is
convincing evidence for antimicrobial secondary metabolites
in this pathogen, excluding that obtained from commonly
observed phytochemicals in the plants. Herein, we aimed to
demonstrate an efficient analytical approach for the production
of antibacterial secondary metabolites using the mycelial
extract of G. boninense. Three experimental cultures were
prepared from fruiting bodies (GBFB), mycelium cultured
on potato dextrose agar (PDA) media (GBMA), and liquid
broth (GBMB). Through solvent extraction, culture type-dependent
phytochemical distributions were diversely exhibited.
Water-extracted GBMB produced the highest yield (31.21
± 0.61%, p < 0.05), but both GBFB and GBMA elicited remarkably
higher yields than GBMB when polar-organic solvent
extraction was employed. Greater quantities of phytochemicals
were also obtained from GBFB and GBMA, in sharp
contrast to those gleaned from GBMB. However, the highest
antibacterial activity was observed in chloroform-extracted
GBMA against all tested bacteria. From liquid-liquid extractions
(LLE), it was seen that mycelia extraction with combined
chloroform-methanol-water at a ratio of 1:1:1 was superior
at detecting antibacterial activities with the most significant
quantities of antibacterial compounds. The data demonstrate
a novel means of assessing antibacterial compounds with mycelia
by LLE which avoids the shortcomings of standardized
method
ologies. Additionally, the antibacterial extract from
the mycelia demonstrate that previously unknown bioactive
secondary metabolites of the less studied subsets of Ganoderma
may serve as active and potent antimicrobial compounds.
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Citations
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