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Título:
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Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
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Autores:
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Torres Bacete, Jesús ;
Hormigo, Daniel ;
Torres Guzmán, Raquel ;
Arroyo Sánchez, Miguel ;
Castillón, María Pilar ;
García, José Luis ;
Acebal Sarabia, Carmen ;
Mata Riesco, Isabel de la
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Tipo de documento:
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texto impreso
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Editorial:
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ASM, 2015-02
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Dimensiones:
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application/pdf
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Nota general:
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cc_by
info:eu-repo/semantics/openAccess
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Idiomas:
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Palabras clave:
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Estado = Publicado
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Materia = Ciencias Biomédicas: Biología
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Materia = Ciencias Biomédicas: Biología: Biología molecular
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Materia = Ciencias Biomédicas: Biología: Microbiología
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Tipo = Artículo
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Resumen:
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The pva gene from Streptomyces lavendulae ATCC 13664, encoding a novel penicillin V acylase (SlPVA), has been isolated and characterized. The gene encodes an inactive precursor protein containing a secretion signal peptide that is activated by two internal autoproteolytic cleavages that release a 25-amino-acid linker peptide and two large domains of 18.79 kDa (_-subunit) and 60.09 kDa (_-subunit). Based on sequence alignments and the three-dimensional model of SlPVA, the enzyme contains a hydrophobic pocket involved in catalytic activity, including Ser_1, His_23, Val_70, and Asn_272, which were confirmed by site-directed mutagenesis studies. The heterologous expression of pva in S. lividans led to the production of an extracellularly homogeneous heterodimeric enzyme at a 5-fold higher concentration (959 IU/liter) than in the original host and in a considerably shorter time. According to the catalytic properties of SlPVA, the enzyme must be classified as a new member of the Ntn-hydrolase superfamily, which belongs to a novel subfamily of acylases that recognize substrates with long hydrophobic acyl chains and have biotechnological applications in semisynthetic antifungal production.
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En línea:
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https://eprints.ucm.es/43288/1/Acebal.%202015.%20Overexpression%20of%20penicillin%20V%20acylase%20from%20Streptomyces%20lavendulae%20and%20elucidation%20of%20its%20catalytic%20residues.pdf
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