A separate family of enzymes within the metallo‐β‐lactamase fold comprises several important proteins acting on nucleic acid substrates, involved in DNA repair (Artemis, SNM1 and PSO2) and RNA processing [cleavage and polyadenylation specificity factor (CPSF) subunit]. Proteins of this family, named β‐CASP after the names of its representative members, possess specific features relative to those of other metallo‐β‐lactamases, that are concentrated in the C‐terminal part of the domain. In this study, using sensitive methods of sequence analysis, we identified highly conserved amino acids specific to the β‐CASP family, some of which were unidentified to date, that are predicted to play critical roles in the enzymatic function. The identification and characterisation of all the extant, detectable β‐CASP members within sequence databases and genome data also allowed us to unravel particular sequence features which are likely to be involved in substrate specificity, as well as to describe new but as yet uncharacterised members which may play critical roles in DNA and RNA metabolism.