Torres Bacete, JesúsHormigo, DanielTorres Guzmán, RaquelArroyo Sánchez, MiguelCastillón, María PilarGarcía, José LuisAcebal Sarabia, CarmenDe La Mata Riesco, Mª Isabel2023-06-182023-06-182015-020099-224010.1128/AEM.02352-14https://hdl.handle.net/20.500.14352/23383The 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.engAtribución 3.0 Españahttps://creativecommons.org/licenses/by/3.0/es/journal articlehttp://aem.asm.org/open access577.2579577.15MicrobiologyPenicillin V AcylaseStreptomyces lavendulaeBiologíaBiología molecular (Biología)Microbiología (Biología)24 Ciencias de la Vida2415 Biología Molecular2414 Microbiología