RT Journal Article
T1 Overexpression of Penicillin V Acylase from Streptomyces lavendulae and Elucidation of Its Catalytic Residues
A1 Torres Bacete, Jesús
A1 Hormigo, Daniel
A1 Torres Guzmán, Raquel
A1 Arroyo Sánchez, Miguel
A1 Castillón, María Pilar
A1 García, José Luis
A1 Acebal Sarabia, Carmen
A1 De La Mata Riesco, Mª Isabel
AB 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.
PB ASM
SN 0099-2240
YR 2015
FD 2015-02
LK https://hdl.handle.net/20.500.14352/23383
UL https://hdl.handle.net/20.500.14352/23383
LA eng
NO Ministerio de Ciencia e Innovación (MICINN)
NO Ministerio de Educación y Ciencia
DS Docta Complutense
RD 12 may 2025