, a group of multi-drug resistant, Gram-positive, aerobic, and partially acid-fast bacteria, are emerging causes of bacterial conjunctivitis and keratitis. However, the pathogenesis of keratitis is largely unknown. To address this, we used New Zealand White rabbits to develop the first eye infection model and conducted tests to study the pathogenesis mechanisms of . There is increasing evidence that biofilms play a significant role in ocular infections, leading us to hypothesize that biofilm formation is crucial for effective infection. In order to look for potential candidate genes which are important in biofilm formation and keratitis. We performed genome sequencing of two ocular isolates, -PW1004 and -PW899, to identify potential virulence factors. Through and studies, we characterized their biological roles in mediating keratitis. Our findings confirmed that is an ocular pathogen by fulfilling Koch's postulates, and using genome sequence data, we identified encoding a mycolyltransferase, as a crucial gene in biofilm formation and causing keratitis in the rabbit model. This is the first report demonstrating the novel role of mycolyltransferase in causing ocular infections. Overall, our findings contribute to a better understanding of pathogenesis and provide a potential target for treatment. Specific inhibitors targeting TmytC could serve as an effective treatment option for infections.