Integrative interpretation of linoleic acid biotransformation by probiotic Pediococcus acidilactici BCB1H via metabolite profiling and enzyme interaction analysis

This study explores the metabolic potential of Pediococcus acidilactici BCBH1 in LA biotransformation using a multidisciplinary approach involving metabolite profiling, bioinformatics tools (Phyre2, InterProScan), molecular docking, and dynamic simulations. The strain exhibited exceptional tolerance to LA, maintaining consistent growth across concentrations from 1 % to 10 % (w/v), surpassing reported limits for other LAB strains. Metabolite profiling revealed 13 distinct compounds, including linoleic acid ethyl ester (19.18 mg/L) and cis-vaccenic acid, highlighting the strain's efficiency in detoxifying LA and synthesizing bioactive lipids. Sequence similarity analysis identified six hypothetical proteins, with three enzymes L-lactate dehydrogenase, glucose-6-phosphate isomerase, and ribose-phosphate pyrophosphokinase structurally and functionally annotated. These enzymes were found to play critical roles in metabolic pathways, including redox balance, energy optimization, and nucleotide biosynthesis. Molecular docking revealed stable binding interactions, with L-lactate dehydrogenase exhibiting a binding energy of −8.9 kJ/mol, glucose-6-phosphate isomerase at −7.6 kJ/mol, and ribose-phosphate pyrophosphokinase at −9.2 kJ/mol. Dynamic simulations confirmed the stability of the protein-ligand complexes, with L-lactate dehydrogenase displaying consistent RMSD values of 2.0 Å and interaction energies dominated by electrostatic and van der Waals forces. This integrative study elucidates the enzymatic and metabolic pathways of LA biotransformation in P. acidilactici BCBH1, emphasizing its potential in functional food production, metabolic engineering, and therapeutic applications. • Pediococcus acidilactici BCB1H showed high LA tolerance and biotransformation. • Identified 13 bioactive metabolites, including LA ethyl ester (19.18 mg/L). •Discovered three novel enzymes with stable ligand interactions. •Insights support probiotic applications in functional foods and metabolic engineering.