Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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HPLC, NMR Based Characterization, Antioxidant and Anticancer Activities of Chemical Constituents from Therapeutically Active Fungal Endophytes

  • Waqas Hussain Shah (Department of Biotechnology, COMSATS University Islamabad, Abbottabad Campus ) ;
  • Wajiha Khan (Department of Biotechnology, COMSATS University Islamabad, Abbottabad Campus ) ;
  • Sobia Nisa (Department of Microbiology, The University of Haripur) ;
  • Michael H.J. Barfuss (Department of Botany and Biodiversity Research, University of Vienna) ;
  • Johann Schinnerl (Department of Botany and Biodiversity Research, University of Vienna) ;
  • Markus Bacher (Department of Chemistry, Institute of Chemistry of Renewable Resources, University of Natural Resources and Life Sciences (BOKU)) ;
  • Karin Valant-Vetschera (Department of Botany and Biodiversity Research, University of Vienna) ;
  • Ashraf Ali (Department of Chemistry, Faculty of Physical and Applied Sciences, The University of Haripur) ;
  • Hiba-Allah Nafidi (Department of Food Science, Faculty of Agricultural and Food Sciences, Laval University) ;
  • Yousef A. Bin Jardan (Department of Pharmaceutics, College of Pharmacy, King Saud University) ;
  • John P. Giesy (Toxicology Centre, University of Saskatchewan)
  • Received : 2024.03.18
  • Accepted : 2024.04.29
  • Published : 2024.07.28
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Abstract

Fungi generate different metabolites some of which are intrinsically bioactive and could therefore serve as templates for drug development. In the current study, six endophytic fungi namely Aspergillus flavus, Aspergillus tubigenesis, Aspergillus oryzae, Penicillium oxalicum, Aspergillus niger, and Aspergillus brasiliensis were isolated and identified from the medicinal plant, Silybum marianum. These endophytic fungi were identified through intra transcribed sequence (ITS) gene sequencing. The bioactive potentials of fungal extracts were investigated using several bioassays such as antibacterial activity by well-diffusion, MIC, MBC, anti-biofilm, antioxidant, and haemolysis. The Pseudomonas aeruginosa PAO1 was used to determine the antibiofilm activity. The ethyl acetate extract of Aspergillus flavus showed strong to moderate efficacy against Staphylococcus aureus, Escherichia coli, P. aeruginosa, and Bacillus spizizenii. Aspergillus flavus and Aspergillus brasiliensis exhibited significant antibiofilm activity with IC50 at 4.02 and 3.63 mg/ml, while A. flavus exhibited maximum antioxidant activity of 50.8%. Based on HPLC, LC-MS, and NMR experiments kojic acid (1) and carbamic acid (methylene-4, 1-phenylene) bis-dimethyl ester (2) were identified from A. flavus. Kojic acid exhibited DPPH free radical scavenging activity with an IC50 value of 99.3 ㎍/ml and moderate activity against ovarian teratocarcinoma (CH1), colon carcinoma (SW480), and non-small cell lung cancer (A549) cell lines. These findings suggest that endophytic fungi are able to produce promising bioactive compounds which deserve further investigation.

Keywords

Acknowledgement

We acknowledge the NMR Centre and MS Centre, Faculty of Chemistry, University of Vienna for measuring NMR and MS spectra. The authors would like to extend their sincere appreciation to the Researchers Supporting Project, King Saud University, Riyadh, Saudi Arabia for funding this work through the project number (RSP2024R457) and Higher education commission (HEC) of Pakistan for the financial support through IRSIP

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