*3.2. Genome Data Mining and Annotation of CYP128 P450s*

CYP128 P450s mining in different bacterial species was carried out following the method described elsewhere [22,31]. Briefly, BLAST analysis was performed with the *M. tuberculosis* H37Rv CYP128A1 (*Rv2268c*) P450 sequence with default settings against individual mycobacterial species genomes at the IMG/M database. Considering the International Cytochrome P450 Nomenclature criteria, i.e., P450s showing >40% identity belong to the same family [43–45], all the hit proteins with more than 40% identity were selected and subjected to P450 characteristic motifs analysis as described elsewhere [34,46,47]. Proteins that showed all P450 characteristic motifs were selected for further analysis. Proteins that were short in amino acid length and had none or only one of the highly conserved P450 motifs, such as EXXR and CXG, were considered P450 fragments and not included in the study. The selected proteins were then subjected to BLAST analysis at the P450 webpage (http://www.p450.unizulu.ac.za/) to identify the named homology protein, in this case CYP128 P450. Hit proteins that showed homology to CYP128 were then selected and different subfamilies were assigned following the International Cytochrome P450 Nomenclature criteria, i.e., P450s showing >55% identity belong to the same subfamily [43–45].
