**3.1 Phylogenetic analysis of KARI from pathogens**

According to the results of the BLAST search with the sequence of KARI protein from *Neisseria gonorrhoeae* FA19 (*Ng* KARI), KARI sequences for human pathogens were obtained. A phylogenetic analysis of the amino acid sequence with reported pathogens was conducted. As shown in the result, *Ng* KARI is closely related to the KARI from the ESKAPE pathogens. All of them are clustered in respective clades (**Figure 1a**).

The sequence alignment of KARI protein from other human pathogens is reported in **Figure 1b**. All tested pathogens share a common ancestor. However, KARI from *S. pneumonia*, *N. meningitidis*, *S. enterica*, *M. tuberculosis*, *B. cereus* and *B. anthracis* belong to a different clade than the other pathogens (*Shigella dysenteriae*, *Shigella flexneri*, *Shigella sonnei*, *Yersinia pseudotuberculosis*, *Yersinia enterocoliticia*, and *Francisella pneumoniae*).

The presence of KARI was also assessed in various members of the gut microbiota, to understand if the latter will be inhibited by KARI inhibitors (**Figure 2**). KARI from *P. dentalis* and *B. fragilis* belong to the same clade, different from the other Gut bacteria (*F. nucleatum*, *B. bifidum*, *Lactobacillus* sp., and *A. muciniphila*).
