Most metagenomic vanT calls are not vancomycin resistance genes

Metagenomic resistome surveys frequently report vanT _G as the dominant glycopeptide resistance gene, but these calls rely on homology without verifying catalytic residues or operon context. By analyzing 107,008 metagenome-assembled genomes, we demonstrate that only 3.7% of predicted vanT _G genes possess the required active-site substitution, with the vast majority being misannotated alanine racemases. Incorporating structural and genomic context corrections reveals a novel split-architecture lineage in gut-associated Erysipelotrichales missed by current profiling tools.