title: The signature amidase from Sulfolobus solfataricus belongs to the CX3C subgroup of enzymes cleaving both amides and nitriles. Ser195 and Cys145 are predicted to be the active site nucleophiles.
abstract: The signature amidase from the extremophile archeum Sulfolobus solfataricus is an enantioselective enzyme that cleaves S-amides. We report here that this enzyme also converts nitriles in the corresponding organic acid, similarly to the well characterized amidase from Rhodococcus rhodochrous J1. The archaeal and rhodococcal enzymes belong to the signature amidases and contain the typical serine-glycine rich motif. They work at different optimal temperature, share a high sequence similarity and both contain an additional CX3C motif. To explain their dual specificity, we built a 3D model of the structure of the S. solfataricus enzyme, which suggests that, in addition to the classical catalytic Ser-cisSer-Lys, a putative additional Cys-cisSer-Lys catalytic site, likely to be responsible for nitrile hydrolysis, is present in these proteins. The results of random and site-directed mutagenesis experiments, as well as inhibition studies support our hypothesis.
authors.affiliation: Centre of Biotechnology-Bioprogress, Anagni, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Cilia
authors.forename: Armando
authors.initials: A
authors.lastname: Fabbri
authors.forename: Monica
authors.initials: M
authors.lastname: Uriani
authors.forename: Giuseppe G
authors.initials: GG
authors.lastname: Scialdone
authors.forename: Sergio
authors.initials: S
authors.lastname: Ammendola
mesh_terms: D000577:Amides / Q000378:metabolism*; D000581:Amidohydrolases / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001665:Binding Sites; D020134:Catalytic Domain; D003545:Cysteine; D008958:Models, Molecular; D009154:Mutation; D009570:Nitriles / Q000378:metabolism*; D011487:Protein Conformation; D012694:Serine; D048229:Sulfolobus solfataricus / Q000201:enzymology*; D013696:Temperature
chemical_list: D000577:Amides; D009570:Nitriles; D012694:Serine; D000581:Amidohydrolases; C019665:amidase; D003545:Cysteine