In-vitro Antimicrobial Screening, Enzymatic Assays and Metabolome Profiling and Insilico Analysis for Anticancer Potency of Talaromyces pinophilus (AVK3), Isolated from the Rhizome of Zingiber officinale

Introduction: Endophytic fungi are prolific sources of bioactive constituents with considerable therapeutic efficiency against various diseases, including cancer, by acting as anticancer and antimicrobial agents. The bioprospecting of novel endophytic secondary metabolites has gained momentum in the treatment of cancer with minimal side-effects. Talaromyces pinophilus (AVK3) is known to produce pinophilin and talaromycesins (A and B), which inhibit cancer cell growth. Aim: To evaluate the isolation, antimicrobial screening, enzymatic assays, and metabolomic profiling (GC-MS analysis) of an endophytic fungus of ginger, Talaromyces pinophilus (AVK3), along with in-silico studies of its anticancer properties. Materials and Methods: An in-vitro experimental study was conducted from June 2023 to May 2024 at Acharya Nagarjuna University, Guntur, Andhra Pradesh, India. Thirty fresh ginger rhizomes were collected from Mamillapalli village, Andhra Pradesh, following strict inclusion criteria (healthy, undamaged rhizomes harvested within 24 hours) and used for the isolation of endophytes. The endophyte AVK3 was isolated using surface sterilisation methods and identified as Talaromyces pinophilus through 18S ribosomal Ribonucleic Acid (rRNA) sequencing (GenBank accession: PP957929). Antimicrobial screening was performed against human pathogens using agar well diffusion and Minimum Inhibitory Concentration (MIC) methods. Enzymatic assays and metabolomic analysis were conducted to identify potential bioactive compounds. The anticancer potential of bioactive compounds was further evaluated through molecular docking studies against cancer target proteins. Results: Talaromyces pinophilus exhibited a high zone of inhibition (30 mm in diameter) against Salmonella typhimurium, with a MIC value of 6.25 µg/mL and an Half maximal Inhibitory Concentration (IC50) of 13.397 µg/mL, demonstrating efficient antibacterial activity. Metabolomic profiling through Gas Chromatography–Mass Spectrometry (GC-MS) analysis revealed bioactive compounds like 3,5-dihydroxy-6-methyl-2,3- dihydro-4H-pyran-4-one; 1,2,3-propanetriol; HMF; 3-acetoxy3-hydroxy-propionic acid methyl ester; 2-piperidinone; furandimethanol; hexahydropyrrolo(1,2-a)pyrazine-1,4-dione; terrain; and 3,6-diisobutyl-2,5-piperazinedione. Enzymatic assays showed the presence of hydrolytic enzymes such as amylase, lipase catalse, gelatinase, and lipases. Conclusion: In-vitro antimicrobial studies and in-silico analysis of anticancer potential demonstrated binding affinities of –19.5 kcal/mol against the lung cancer target protein (PDB ID: 7W8O) and –22.9 kcal/mol against the breast cancer target protein (PDB ID: 3HBS), confirming Talaromyces pinophilus as a promising source for anticancer treatment. The extensive array of bioactive compounds exemplifies the chemical diversity of Talaromyces pinophilus, particularly highlighting its potential in biomedical innovation.