Fig. 3

Sequence conservation of the FAD lid and the activation loop and architecture of the active site of OhySa, OhyEm, and OhyRe. A Amino acid sequence alignment of OhyRe (Uniprot: T5I9M6), OhySa (Uniprot: A0A0D6GJV1), OhyLa (Uniprot: Q5FL96), OhyEm (Uniprot: OLHYD), OhySt (Uniprot: A0A126NKL7) restricted to the FAD-lid and the activation loop. Conserved residues in lid and activation loop are highlighted by yellow background. The catalytic residue in the loop is highlighted with a light blue or light orange box, respectively. Highly conserved residues are indicated with asterisk, moderate conservation with two points, low conservation with one point. Primary sequences of Ohys were aligned using Clustal Omega [60]. B Active site of OhyEm (PDB-ID 4uir; [46]), shown with catalytic important residues. The bound PEG molecule in close proximity of the active site is shown in orange. Structural elements shown in cartoon representation. C Active site of OhySa (PDB-ID: 7kaz; [47]) shown with important residues lining the active site. The ternary complex of OhySa with bound FAD and oleate was obtained with the OhySa variant E82A. For clarity, we have computationally re-introduced the wild-type situation. FAD, oleic acid and indicated residues shown in stick representation. D Active site of OhyRe (PDB-ID: 5odo; [48]) shown with important residues lining the active site in stick representation. The shown FAD cofactor and oleic acid were obtained by a superposition of the OhySa structure A and derived from the superposition with the structure of OhySa. E Superposition of the active site of OhyRe and OhySa