Conservation analyses
The PETase-profile HMM was applied for a standard numbering scheme, by aligning the 2930 sequences of PETase homologues from the LED against the respective profile HMM and subsequently assigning the position numbers from the Is PETase reference sequence as standard position numbers. For conservation analysis of PETase homologues, the frequency of amino acid residues or gaps was counted at each standard position.
For the conservation analysis of PURase homologues from LED superfamilies 11 and 13, two multiple sequence alignments were generated using Clustal Omega (version 1.2.4) 23, and the frequency of amino acid residues or gaps was counted at selected positions.