Khattak Saeed Ullah, Iqbal Zafar, Lutfullah Ghosia, Ahmad Sajjad, Alharbi Metab, Alasmari Abdullah F, Irfan Muhammad
Center of Biotechnology and Microbiology, University of Peshawar, KPK, Pakistan.
Department of Agricultural Chemistry, University of Agriculture, KPK, Pakistan.
J Biomol Struct Dyn. 2024;42(23):12776-12787. doi: 10.1080/07391102.2023.2273435. Epub 2023 Oct 25.
Following preliminary bioactivity testing, the fungal strain identified as 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, 2, 3, 4-tetrahydroquinolin-6-yl) vinyl) cyclohexanone with the chemical formula CHNO 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 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) 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 experiments were encouraging as at concentrations of 500 and 1000 μg mL the compound inhibited the growth of by 17% and 38% while 19% and 41% inhibition were recorded against . The compound showed antifungal activity and against both the species and can act as a potent antifungal candidate in the future upon further investigation.Communicated by Ramaswamy H. Sarma.
经过初步生物活性测试后，将鉴定为[具体菌株名称未给出]的真菌菌株在改良的查氏酵母肉汤培养基（CYB）中培养，以生产抗真菌化合物。使用了包括高效液相色谱（HPLC）在内的几种色谱技术从粗提物中纯化真菌代谢产物。使用沃特世液相色谱 - 质谱联用系统对纯化后的化合物进行质量测定，同时使用400兆赫和500兆赫的瓦里安核磁共振仪阐明该化合物的结构。纯化后化合物的化学名称为(2 R, 4S) -2, 4 - 二甲基 - 4 - ((E) - 2 - ((3S, 4S) - 2, 4, 5 - 三羟基 - 3 - 甲氧基 - 4 - 苯基, 2, 3, 4 - 四氢喹啉 - 6 - 基)乙烯基)环己酮，化学式为C₂₅H₃₅NO₅，精确质量为437.2。分子对接预测该化合物与二氢叶酸还原酶和羊毛甾醇14α - 脱甲基酶的对接分数分别为 - 8.1千卡/摩尔和 - 9.8千卡/摩尔。此外，该化合物与这些酶显示出稳定的结合模式，并具有较强的结合能。经过计算机模拟分析后，使用琼脂管扩散法对质量为437的该化合物针对两种致病性酵母菌种（即白色念珠菌和光滑念珠菌）的抗真菌潜力进行了测试。使用无菌二甲基亚砜（DMSO）将该化合物配制成四种剂量浓度（100、250、500、1000微克/毫升），并与高压灭菌的半固体马铃薯葡萄糖琼脂（PDA）培养基在标有相应剂量浓度的螺口试管中混合。将真菌菌株接种在这种培养基上，并以氟康唑作为对照药物计算真菌菌株的线性生长抑制率。实验结果令人鼓舞，在500和1000微克/毫升的浓度下，该化合物对白色念珠菌的生长抑制率分别为17%和38%，对光滑念珠菌的抑制率分别为19%和41%。该化合物对两种念珠菌属菌种均显示出抗真菌活性，经进一步研究后未来可作为一种有效的抗真菌候选药物。由拉马斯瓦米·H·萨尔马传达。
