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dc.contributor.authorSoriano Maldonado, Pablo 
dc.contributor.authorAndújar Sánchez, Montserrat 
dc.contributor.authorClemente Jiménez, Josefa María 
dc.contributor.authorRodríguez Vico, Felipe 
dc.contributor.authorLas Heras Vázquez, Francisco Javier 
dc.contributor.authorMartínez Rodríguez, Sergio 
dc.date.accessioned2024-05-22T08:48:51Z
dc.date.available2024-05-22T08:48:51Z
dc.date.issued2015
dc.identifier.issn1559-0305
dc.identifier.urihttp://hdl.handle.net/10835/16503
dc.description.abstractN-Succinyl-amino acid racemase (NSAAR), long referred to as N-acyl- or N-acetyl-amino acid racemase, is an enolase superfamily member whose biotechnological potential was discovered decades ago, due to its use in the industrial dynamic kinetic resolution methodology first known as ‘‘Acylase Process’’. In previous works, an extended and enhanced substrate spectrum of the NSAAR from Geobacillus kaustophilus CECT4264 toward different N-substituted amino acids was reported. In this work, we describe the cloning, purification, and characterization of the NSAAR from Geobacillus stearothermophilus CECT49 (GstNSAAR). The enzyme has been extensively characterized, showing a higher preference toward N-formyl-amino acids than to N-acetyl-amino acids, thus confirming that the use of the former substrates is more appropriate for a biotechnological application of the enzyme. The enzyme showed an apparent thermal denaturation midpoint of 77.0 ± 0.1 C and an apparent molecular mass of 184 ± 5 kDa, suggesting a tetrameric species. Optimal parameters for the enzyme activity were pH 8.0 and 55–65 C, with Co2? as the most effective cofactor. Mutagenesis and binding experiments confirmed K166, D191, E216, D241, and K265 as key residues in the activity of GstNSAAR, but not indispensable for substrate binding.es_ES
dc.language.isoenes_ES
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 Internacional*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectN-Succinyl-amino acid racemasees_ES
dc.subjectN-Acylamino acid racemasees_ES
dc.subjectN-Acetyl-amino acid racemasees_ES
dc.subjectAcylase processes_ES
dc.subjectAmino acides_ES
dc.subjectRacemasees_ES
dc.titleBiochemical and Mutational Characterization of N-Succinyl- Amino Acid Racemase from Geobacillus stearothermophilus CECT49es_ES
dc.typeinfo:eu-repo/semantics/articlees_ES
dc.rights.accessRightsinfo:eu-repo/semantics/openAccesses_ES


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Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivatives 4.0 Internacional