Multiple amino acid sequence alignments and phylogenetic analysis of EstCS2. (a) Multiple amino acid sequence alignments of EstCS2 and its homologs. ZP_03881667, carboxylesterase from Haliangium ochraceum DSM 14365; ZP_04761385, Carboxylesterase from Acidovorax delafieldii 2AN; BAA76305, polyurethane esterase from Delftia acidovorans YP_001136305, carboxylesterase from Mycobacterium gilvum PYR-GCK; ZP_03620470, Carboxylesterase from Tolumonas auensis DSM 9187 AAL82802, paraben-hydrolyzing esterase precursor from Enterobacter cloacae; YP_001069337, carboxylesterase, from Mycobacterium sp. JLS. (b) Phylogenetic tree based on amino acid sequence of EstCS2 and closely related proteins. Phylogenetic analysis was performed using the program MEGALIGN (DNASTAR, Madison, WI). Except EstCS2, protein sequences for previously identified families of bacterial lipolytic enzymes were retrieved from GenBank (http://www.ncbi.nlm.nih.gov). The units at the bottom of the tree indicate the number of substitution events.