title: N-Carbamoyl-β-alanine amidohydrolase from Agrobacterium tumefaciens C58: a promiscuous enzyme for the production of amino acids.
abstract: The availability of enzymes with a high promiscuity/specificity relationship permits the hydrolysis of several substrates with a view to obtaining a certain product or using one enzyme for several productive lines. N-Carbamoyl-β-alanine amidohydrolase from Agrobacterium tumefaciens (Atβcar) has shown high versatility to hydrolyze different N-carbamoyl-, N-acetyl- and N-formyl-amino acids to produce different α, β, γ and δ amino acids. We have calculated the promiscuity index for the enzyme, obtaining a value of 0.54, which indicates that it is a modestly promiscuous enzyme. Atβcar presented the highest probability of hydrolysis for N-carbamoyl-amino acids, being the enzyme more efficient for the production of α-amino acids. We have also demonstrated by mutagenesis, modelling, kinetic and binding experiments that W218 and A359 indirectly influence the plasticity of the enzyme due to interaction with the environment of R291, the key residue for catalytic activity.
authors.affiliation: Dpto. Química-Física, Bioquímica y Química Inorgánica, Universidad de Almería Edificio CITE I, Carretera de Sacramento s/n, 04120 Almería, Spain.
authors.forename: A I
authors.initials: AI
authors.lastname: Martínez-Gómez
authors.forename: M
authors.initials: M
authors.lastname: Andújar-Sánchez
authors.forename: J M
authors.initials: JM
authors.lastname: Clemente-Jiménez
authors.forename: J L
authors.initials: JL
authors.lastname: Neira
authors.forename: F
authors.initials: F
authors.lastname: Rodríguez-Vico
authors.forename: S
authors.initials: S
authors.lastname: Martínez-Rodríguez
authors.forename: F J
authors.initials: FJ
authors.lastname: Las Heras-Vázquez
mesh_terms: D016960:Agrobacterium tumefaciens / Q000201:enzymology* / Q000235:genetics; D000581:Amidohydrolases / Q000737:chemistry* / Q000378:metabolism*; D000596:Amino Acids / Q000096:biosynthesis*; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004548:Elasticity; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity; D013816:Thermodynamics; D015091:beta-Alanine / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D011994:Recombinant Proteins; D015091:beta-Alanine; D000581:Amidohydrolases; C451299:N-carbamoylamino acid amidohydrolase