Conjugated linoleic acids (CLAs) are bioactive lipid isomers with potential health-promoting properties. Certain gut microbes, especially members of Lactobacillus genera, can convert linoleic acid (LA) into CLAs through specialized metabolic pathways. However, the genetic basis and functional diversity of CLA biosynthesis in gut microbial communities remain incompletely understood. To investigate the genomic potential for CLA production in gut-associated Lactobacillus taxa, we plan first to use two isolated Lactobacillus strains to identify genes encoding CLA-converting enzymes, along with their 200 bp upstream and downstream flanking sequences. Gene knockout technology is then applied to evaluate the functional roles of these genes. Subsequently, we will retrieve all available whole-genome sequencing (WGS) data and isolated Lactobacillus strains from NCBI to catalog the diversity of enzyme variants involved in CLA bioconversion. In the next step, we will further collect all Lactobacillus genomes, including those derived from metagenomic and shotgun sequencing datasets, and perform HMM-based searches to identify CLA isomerase homologs across gut microbial genomes. Finally, we assess the abundance of Lactobacillus and its corresponding CLA isomerase genes in cohorts of healthy and diabetic individuals (10,000) to explore potential differences and associations.