Supercritical CO₂-extracted oils from Trachyspermum ammi (carom) seeds: Transitioning from food relevance to antifungal mechanisms against Aspergillus fumigatus

• Trachyspermum ammi seeds oil (TaSO) combats A. fumigatus growth. TaSO exhibits anti-melanogenic effect via downregulation of PksP . • TaSO suppresses ergosterol biosynthesis by regulating Erg1, Erg6 , and SrbA, AtrA . • TaSO inhibits biofilm formation and eradicated matured A. fumigatus biofilms. • Thymol orchestrates volatile anti- Aspergillus activity of TaSO. • Petroselinic acid in TaSO also exhibits anti- Aspergillus properties. Traditionally, a kitchen’s toolkit for enhancing flavors, culinary spices stand out as repertoire of bioactive phytoconstituents with robust anti-microbial activity. Aspergillus fumigatus is an opportunistic fungal pathogen responsible for wide spectrum of infections. With the extensive use of fungicides, there has been a surge in multi-drug-resistant A. fumigatus strains. The current study uncovers the anti- Aspergillus effects of Supercritical-CO 2 -extracted oil from Trachyspermum ammi (L.) Sprague (carom) seeds (hereafter, TaSO). TaSO impairs metabolic activity and suppresses growth, hyphal development, biomass accumulation and conidial germination in A. fumigatus . In disc diffusion assay, TaSO produces a clear inhibitory zone with a prominent demelanization effect against A. fumigatus . TaSO exhibits anti-melanogenic effect by PksP downregulation; and attenuates total ergosterol levels in A. fumigatus with correlational downregulation of genes regulating ergosterol biosynthesis, Erg1, Erg6 and transcriptional regulators, SrbA and AtrA . TaSO prevents A. fumigatus biofilm formation and concurrently disrupts pre-formed mature biofilms. GC-MS/MS analysis reveals thymol as major volatile component of TaSO along with fatty acids, petroselinic acid (PetFA), palmitic acid and linoleic acid. Notably, the volatile components of TaSO predominantly driven by Thymol markedly suppresses the growth of A. fumigatus , without being in direct contact. Whereas both Thymol and PetFA, in-contact, independently mediates demelanization and growth inhibitory effects against A. fumigatus by downregulating PksP . Taken together, the observed multifaceted antifungal action of TaSO against A. fumigatus substantiates the age-old belief, Food as Medicine . The current study emphasizes on bridging the food-medicine interface that integrates nutrition, safety and preventive health against fungal diseases.