Ph.D. Defence of M. Atif Khan
CONDUCTION OF PUBLIC DEFENCE IN RESPECT OF MR. MUHAMMAD ATIF KHAN, PH. D RESEARCH SCHOLAR, CENTRE OF BIOTECHNOLOGY AND MICROBIOLOGY, UNIVERISTY OF PESHAWAR
Date: 21-04-2026
Time: 11:30 a.m
Venue: Video Conference Hall
Abstract
Microorganisms hold tremendous potential as a vast reservoir of natural. Products. They serve as a prolific source. of bioactive. Metabolites with diverse applications, including but not limited to agrochemicals, antiparasitic, antibiotics, immunosuppressants, and anticancer agents. In present investigation we selected two fungal species Trichoderma koningii (T. koningii) and Penicillium herquei (P. herquei) to explore their biological potential and production of secondary metabolites. Optimization was done by different growth parameters (incubation period, temperature, nutrient media, pH and (static/shaking intervals), so from both fungal species maximum production of secondary metabolites were achieved.
Five different nutritional media were used in present study, for both P. herquei and T. koningii maximum production of metabolites were observed in Potato Dextrose Broth (PDB).
Large scale fermentation of both fungi T. koningii and P. herquei was done in PDB, extracted with organic solvent Ethyl acetate (EtOAc) and fractionated with n- hexane. The EtOAc fractions of both fungal species were subjected to thin. layer.chromatography,.Column chromatography. and.Preparative thin-layer chromatography, which yielded seven different known compounds. The identification of these isolated compounds was accomplished through a comprehensive analysis utilizing state-of-the-art spectroscopic methods, including 1H-NMR, 13C-NMR, UV, IR. The compounds isolated from EtOAc fraction of T. koningii were 6-Pentyl-2H-pyran-2-one (1) koningin A (2). Chemical constituents extracted from EtOAc fraction of P. herquei were peniciherqueinone (3), linoleic acid (4), kynapcin-24 (5), kojic acid (6) and stigmasterol (7).
Metabolomic profiling of both fungal species were studied by utilizing LCMS-QTOF and GCMS techniques. The GCMS of T. koningii and P. herquei revealed the presence of certain metabolites such as oleic acid, Phenol, 2,4-bis(1,1-dimethylethyl) 2-Nonadecanone 2,4-dinitrophenylhydrazin and Dasycarpidan-1-methanol acetate. The LCMS-QTOF investigations verified the presence of diverse secondary metabolites within the ethyl acetate extract of T. koningii. And P. herquei at different retention time such as Amobarbital, Maculosin, Nigakilactone A and Kurilensoside F. The EtOAc fraction of T. koningii showed significant antibacterial potential against X. campestris. The EtOAc fraction of P. herquei against S. aureus displayed promising antibacterial activity. T. koningii EtOAc against A. solani displayed significant mycological activity, while moderate activity was displayed against other tested fungi. Against F. oxysporum the crude EtOAc and n-hexane extract of P.herquei showed significant activity. T. koningii and P. herquei EtOAc extracts at higher concentration (1000 ugml-1) showed significant inhibitory affect against lemna minor. At higher concentration (1000 ugml-1) EtOAc extract of T. koningii and P. herquei showed high mortality against Callosobruchus analis.
In vivo activities (acute toxicity, anti-inflammatory, antinociceptive, muscle relaxant and antispasmodic) on an animal model (mice) were evaluated. exhibited no sign of acute toxicity was not recorded at a dose level of 10-20 ml/kg by .EtOAc extract of both fungal species throughout the 24h observation. A Significant.decrease in inflammation (p < 0.001) in the carrageenin-induced paw edema test was exhibited by ethyl acetate extract of P. herquei and T. koningii during 5h study. A significant (p < 0.001) percent inhibition of nociception was demonstrated at dose concentration 150 mg/kg by T. koningii and P. herquei. In antispasmodic activity, the EtOAc extract of T. Koningii and P. herquei has significant effect on gut motility compare to caster oil. Although the differences between the doses of P. herquei and T. koningii was nonsignificant. Anticancer studies on cancer lines HeLa, 3T3 and PC3, significant proliferation(percent inhibition) was recorded for P. herquei against PC3 Cell lines at high dose concentration 60ugml-1
Extracellular enzymatic assay was performed on both P. herquei and T. koningii for their evaluation of commercially important enzymes. P. herquei showed production of amylase, cellulose and protease. T. koningii showed production of amylase, cellulase, protease and lipase.
In silico investigation, of tentatively identified compounds from LC-MS-QTOF analysis were conducted to assess their potential interactions with bacterial enzyme tyrosine (Tyrosyl-Trna Synthetase) and an inflammation-supporting enzyme COX-2. These simulations validated the antibacterial and anti-inflammatory activities of the extract from P. herquei. Ligand (L9) had highest binding interaction against both targeted enzymes (1TYD, COX-2). Bacterial protein (1TYD) gave -10.1 Kcalmol-1 binding affinity while against COX-2 it showed -10.2 Kcalmol-1.
In conclusion significantly potent biological and pharmacological potential was demonstrated by both fungi. Metabolites from T. koningii and P. herquei show strong pharmacological potential as leads for drug development, while their enzymatic activities suggest valuable applications in industrial biotechnology.