TY - JOUR 
A1 - Torres Bacete, Jesús
AU - Hormigo Cisneros, Daniel
AU - Torres Guzmán, Raquel
AU - Arroyo, Miguel
AU - Castillón, María Pilar
AU - García, José Luis
AU - Acebal Sarabia, Carmen
AU - Mata Riesco, Isabel de la
T1 - Overexpression of penicillin V acylase from streptomyces lavendulae and elucidation of its catalytic residues
Y1 - 2015
SN - 00992240

UR - http://hdl.handle.net/11268/4436
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 (α-subunit) and 60.09 kDa (β-subunit). Based on sequence alignments and the three-dimensional model of SlPVA, the enzyme contains a hydrophobic pocket involved in catalytic activity, including Serβ1, Hisβ23, Valβ70, and Asnβ272, 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.
KW - Biotecnología
KW - Enzimas
KW - Biotecnología
KW - Enzima
LA - eng
ER -