title: Promiscuous enantioselective (-)-γ-lactamase activity in the Pseudomonas fluorescens esterase I.
abstract: A promiscuous but very enantioselective (-)-γ-lactamase activity in the kinetic resolution of the Vince lactam (2-azabicyclo[2.2.1]hept-5-en-3-one) was detected in the Pseudomonas fluorescens esterase I (PFEI). The lactamase activity was increased 200-fold by the introduction of a point mutation and resulted as enantioselective as the Microbacterium sp. enzyme used industrially in this resolution. The structural and mechanistic determinants for the catalytic promiscuity and enantioselectivity were identified by molecular modeling, setting a ground stone to engineer further amidase-related activities from this esterase.
authors.affiliation: Center for Molecular Biology Severo Ochoa (UAM-CSIC), Nicolás Cabrera 1, E-28049 Madrid, Spain.
authors.forename: Leticia L
authors.initials: LL
authors.lastname: Torres
authors.forename: Anna
authors.initials: A
authors.lastname: Schliessmann
authors.forename: Marlen
authors.initials: M
authors.lastname: Schmidt
authors.forename: Noella
authors.initials: N
authors.lastname: Silva-Martin
authors.forename: Juan A
authors.initials: JA
authors.lastname: Hermoso
authors.forename: José
authors.initials: J
authors.lastname: Berenguer
authors.forename: Uwe T
authors.initials: UT
authors.lastname: Bornscheuer
authors.forename: Aurelio
authors.initials: A
authors.lastname: Hidalgo
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism*; D043182:Carboxylesterase / Q000737:chemistry / Q000378:metabolism*; D007769:Lactams / Q000737:chemistry / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D011487:Protein Conformation; D011551:Pseudomonas fluorescens / Q000201:enzymology*; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D007769:Lactams; C062350:2-azabicyclo(2.2.1)hept-5-en-3-one; D043182:Carboxylesterase; C469486:Ces1c protein, mouse; D000581:Amidohydrolases; C401943:gamma-lactamase