Recruitment of lysosomal cathepsins B, L and D as digestive enzymes in Coleoptera

The recruitment of the lysosomal cathepsins B (CAB), L (CAL) and D (CAD) as luminal digestive enzymes was investigated in 3 species of beetles. Gene expression was determined by RNA‐seq in different regions of the midgut and in the carcasses from the transcriptomes of Dermestes maculatus, Tenebrio molitor and Zabrotes subfasciatus. These data together with phylogenetic analyses, allowed us to identify the sequences of the gene coding for digestive and lysosomal CABs, CADs and CALs in T. molitor and Z. subfasciatus and observe the absence of digestive cathepsins in D. maculatus. Comparisons of structures based on the overall similarity of modelled structures were performed and subsite residues in the lysosomal and digestive CALs were identified by molecular docking. The data showed that S2 subsites are very variable, probably as an adaption to a luminal digestive role. The survey of sequences of the gene coding for cathepsins in the genomes of 13 beetle species from different phylogenetic groups showed that expansion of CAL and CAB genes occurred only in the Cucujiformia clade. Several digestive CABs have a reduced occluding loop, probably to act as digestive enzymes. Pollen‐feeding was proposed to be the selective pressure to recruit cathepsins as digestive enzymes in Cucujiformia beetles. The recruitment of the cathepsins B, L and D as luminal digestive enzymes was investigated in beetles. The expansion of CAL and CAB genes occurred only in the clade Cucujiformia. Structural modifications in digestive CABs and CALs are described. Pollen feeding is proposed as a possible ancestral diet of herbivorous beetles.