Exploring Antimycobacterial Potential: Safety Evaluation and Active Compound Isolation from <i>Gymnopilus junonius</i>

Background/Objectives: Tuberculosis remains a major public health crisis, and it is imperative to search for new antimycobacterial drugs. Natural products, including medicinal macrofungi, have been used as sources for the discovery of pharmaceuticals; however, research on their antimycobacterial act...

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Main Authors: Jenske Didloff, Gerhardt J. Boukes, Mutenta N. Nyambe, Denzil R. Beukes, Mookho S. Lerata, Velile Vilane, Michael Lee, Sharlene Govender, Maryna van de Venter
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Antibiotics
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Online Access:https://www.mdpi.com/2079-6382/14/2/179
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author Jenske Didloff
Gerhardt J. Boukes
Mutenta N. Nyambe
Denzil R. Beukes
Mookho S. Lerata
Velile Vilane
Michael Lee
Sharlene Govender
Maryna van de Venter
author_facet Jenske Didloff
Gerhardt J. Boukes
Mutenta N. Nyambe
Denzil R. Beukes
Mookho S. Lerata
Velile Vilane
Michael Lee
Sharlene Govender
Maryna van de Venter
author_sort Jenske Didloff
collection DOAJ
description Background/Objectives: Tuberculosis remains a major public health crisis, and it is imperative to search for new antimycobacterial drugs. Natural products, including medicinal macrofungi, have been used as sources for the discovery of pharmaceuticals; however, research on their antimycobacterial activity remains limited. This study aimed to isolate and identify the bioactive compound responsible for antimycobacterial activity, thereby expanding on the limited knowledge regarding the antimicrobial activity and bioactive compounds present in <i>Gymnopilus junonius</i>. Methods: Bioassay-guided fractionation using column chromatography and preparative thin-layer chromatography were employed to isolate the active compound. Antimycobacterial activity against <i>Mycobacterium tuberculosis</i> H37 was assessed using a resazurin microplate assay (REMA). The chemical structure was determined by <sup>1</sup>H nuclear magnetic resonance (NMR) spectroscopy, heteronuclear single quantum coherence (HSQC) spectroscopy, heteronuclear multiple bond correlation (HMBC) spectroscopy, and high-resolution electrospray ionization mass (HR-ESI-MS) spectrometry. Transmission electron microscopy (TEM) was used to observe the ultrastructural changes in <i>M</i>. <i>tuberculosis</i> induced by the compound. Cytotoxicity was evaluated in African green monkey kidney cells (Vero), human liver cells (C3A), and zebrafish embryos/larvae. Results: Bioassay-guided fractionation led to the isolation of gymnopilene, which showed inhibitory activity against <i>M</i>. <i>tuberculosis</i> (MIC: 31.25 µg/mL). TEM analysis revealed that treatment with gymnopilene caused ultrastructural damage observed as the disruption and disintegration of the cell wall. While gymnopilene demonstrated cytotoxicity in Vero and C3A cells, no toxicity was observed in zebrafish embryos/larvae for the crude extract. Conclusions: This study highlights that macrofungi, such as <i>G</i>. <i>junonius</i>, could be a valuable resource of bioactive compounds.
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spelling doaj-art-ea922eb460064127ae14680e15faacf42025-08-20T02:44:55ZengMDPI AGAntibiotics2079-63822025-02-0114217910.3390/antibiotics14020179Exploring Antimycobacterial Potential: Safety Evaluation and Active Compound Isolation from <i>Gymnopilus junonius</i>Jenske Didloff0Gerhardt J. Boukes1Mutenta N. Nyambe2Denzil R. Beukes3Mookho S. Lerata4Velile Vilane5Michael Lee6Sharlene Govender7Maryna van de Venter8Department of Biochemistry and Microbiology, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South AfricaDepartment of Biochemistry and Microbiology, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South AfricaDepartment of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmacy, Nutrition and Dietetics, Lusaka Apex Medical University, Lusaka 10101, ZambiaSchool of Pharmacy, University of Western Cape, Bellville 7535, South AfricaSchool of Pharmacy, University of Western Cape, Bellville 7535, South AfricaCenter for High-Resolution Transmission Electron Microscopy (HRTEM), Physics Department, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South AfricaCenter for High-Resolution Transmission Electron Microscopy (HRTEM), Physics Department, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South AfricaDepartment of Biochemistry and Microbiology, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South AfricaDepartment of Biochemistry and Microbiology, Nelson Mandela University, P.O. Box 77000, Gqeberha 6031, South AfricaBackground/Objectives: Tuberculosis remains a major public health crisis, and it is imperative to search for new antimycobacterial drugs. Natural products, including medicinal macrofungi, have been used as sources for the discovery of pharmaceuticals; however, research on their antimycobacterial activity remains limited. This study aimed to isolate and identify the bioactive compound responsible for antimycobacterial activity, thereby expanding on the limited knowledge regarding the antimicrobial activity and bioactive compounds present in <i>Gymnopilus junonius</i>. Methods: Bioassay-guided fractionation using column chromatography and preparative thin-layer chromatography were employed to isolate the active compound. Antimycobacterial activity against <i>Mycobacterium tuberculosis</i> H37 was assessed using a resazurin microplate assay (REMA). The chemical structure was determined by <sup>1</sup>H nuclear magnetic resonance (NMR) spectroscopy, heteronuclear single quantum coherence (HSQC) spectroscopy, heteronuclear multiple bond correlation (HMBC) spectroscopy, and high-resolution electrospray ionization mass (HR-ESI-MS) spectrometry. Transmission electron microscopy (TEM) was used to observe the ultrastructural changes in <i>M</i>. <i>tuberculosis</i> induced by the compound. Cytotoxicity was evaluated in African green monkey kidney cells (Vero), human liver cells (C3A), and zebrafish embryos/larvae. Results: Bioassay-guided fractionation led to the isolation of gymnopilene, which showed inhibitory activity against <i>M</i>. <i>tuberculosis</i> (MIC: 31.25 µg/mL). TEM analysis revealed that treatment with gymnopilene caused ultrastructural damage observed as the disruption and disintegration of the cell wall. While gymnopilene demonstrated cytotoxicity in Vero and C3A cells, no toxicity was observed in zebrafish embryos/larvae for the crude extract. Conclusions: This study highlights that macrofungi, such as <i>G</i>. <i>junonius</i>, could be a valuable resource of bioactive compounds.https://www.mdpi.com/2079-6382/14/2/179<i>Gymnopilus junonius</i>antimycobacterial activitygymnopilene<i>Mycobacterium tuberculosis</i>cytotoxicity
spellingShingle Jenske Didloff
Gerhardt J. Boukes
Mutenta N. Nyambe
Denzil R. Beukes
Mookho S. Lerata
Velile Vilane
Michael Lee
Sharlene Govender
Maryna van de Venter
Exploring Antimycobacterial Potential: Safety Evaluation and Active Compound Isolation from <i>Gymnopilus junonius</i>
Antibiotics
<i>Gymnopilus junonius</i>
antimycobacterial activity
gymnopilene
<i>Mycobacterium tuberculosis</i>
cytotoxicity
title Exploring Antimycobacterial Potential: Safety Evaluation and Active Compound Isolation from <i>Gymnopilus junonius</i>
title_full Exploring Antimycobacterial Potential: Safety Evaluation and Active Compound Isolation from <i>Gymnopilus junonius</i>
title_fullStr Exploring Antimycobacterial Potential: Safety Evaluation and Active Compound Isolation from <i>Gymnopilus junonius</i>
title_full_unstemmed Exploring Antimycobacterial Potential: Safety Evaluation and Active Compound Isolation from <i>Gymnopilus junonius</i>
title_short Exploring Antimycobacterial Potential: Safety Evaluation and Active Compound Isolation from <i>Gymnopilus junonius</i>
title_sort exploring antimycobacterial potential safety evaluation and active compound isolation from i gymnopilus junonius i
topic <i>Gymnopilus junonius</i>
antimycobacterial activity
gymnopilene
<i>Mycobacterium tuberculosis</i>
cytotoxicity
url https://www.mdpi.com/2079-6382/14/2/179
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