Purification and structure elucidation of Penicillium chrysogenum derived antifungal compound with potential anti-Candida property: in silico and in vitro evidence

Saeed Ullah Khattak; Zafar Iqbal; Ghosia Lutfullah; Sajjad Ahmad; Metab Alharbi; Abdullah F Alasmari; Muhammad Irfan

doi:10.1080/07391102.2023.2273435

Purification and structure elucidation of Penicillium chrysogenum derived antifungal compound with potential anti-Candida property: in silico and in vitro evidence

J Biomol Struct Dyn. 2023 Oct 25:1-12. doi: 10.1080/07391102.2023.2273435. Online ahead of print.

Authors

Saeed Ullah Khattak¹, Zafar Iqbal², Ghosia Lutfullah¹, Sajjad Ahmad^{3

4

5

6}, Metab Alharbi⁷, Abdullah F Alasmari⁷, Muhammad Irfan⁸

Affiliations

¹ Center of Biotechnology and Microbiology, University of Peshawar, KPK, Pakistan.
² Department of Agricultural Chemistry, University of Agriculture, KPK, Pakistan.
³ Department of Health and Biological Sciences, Abasyn University, Peshawar, Pakistan.
⁴ Gilbert and Rose-Marie Chagoury School of Medicine, Lebanese American University, Beirut, Lebanon.
⁵ Department of Natural Sciences, Lebanese American University, Beirut, Lebanon.
⁶ Department of Computer Science, Virginia Tech, USA.
⁷ Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
⁸ Department of Oral Biology, College of Dentistry, University of Florida, Gainesville, FL, USA.

PMID: 37878068
DOI: 10.1080/07391102.2023.2273435

Abstract

Following preliminary bioactivity testing, the fungal strain identified as Penicillium crysogenum was cultured in a modified Czapec Yeast Broth medium (CYB) for the production of antifungal compounds. Several chromatographic techniques including HPLC were used to purify the fungal metabolites from the crude extract. The mass determination of the purified compound was performed using Water's LCMS system while the structure of the compound was elucidated using 400 and 500 Varian NMR machines. The chemical name of the purified compound is (2 R, 4S) -2, 4-dimethyl-4-((E)-2-((3S, 4S)-2, 4, 5-trihydroxy-3-methoxy-4-phenyl^-1, 2, 3, 4-tetrahydroquinolin-6-yl) vinyl) cyclohexanone with the chemical formula C₂₆H₃₁NO₅ and exact mass of 437.2. Molecular docking predicted compound docking score with dihydrofolate reductase enzyme and lanosterol 14α-demethylase enzyme as -8.1 kcal/mol and -9.8 kcal/mol respectively. Further, the compounds showed stable binding mode with the enzymes and reported robust binding energies. After insilico analysis, the compound with mass 437 was tested for its antifungal potential in vitro against two pathogenic yeast species (i.e. Candida albicans and Candida glaberata) using the agar tube diffusion method. Using sterile di-methyl sulfoxide (DMSO) the compound was prepared in four dose concentrations (100, 250, 500, 1000 µg mL^-1) and mixed with autoclaved semisolid Potato Dextrose Agar (PDA) medium in screw-capped test tubes labelled with the corresponding dose concentration. The fungal strains were inoculated on this medium and linear growth inhibition of the fungal strains was calculated using fluconazole as the control drug. The results from in vitro experiments were encouraging as at concentrations of 500 and 1000 μg mL^-1 the compound inhibited the growth of C. albicans by 17% and 38% while 19% and 41% inhibition were recorded against C. glaberata. The compound showed antifungal activity in silico and in vitro against both the Candida species and can act as a potent antifungal candidate in the future upon further investigation.Communicated by Ramaswamy H. Sarma.

Keywords: Candida albicans; Candida glaberata; LCMS; NMR; antifungal compound; insilico.