Computational design of an amidase by combining the best electrostatic features of two promiscuous hydrolases
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Otros documentos de la autoría: Galmes, Miquel Angel; Nödling, Alexander; He, Kaining; Luk, Louis Yu Pan; Świderek, Katarzyna; Moliner, Vicent
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Título
Computational design of an amidase by combining the best electrostatic features of two promiscuous hydrolasesAutoría
Fecha de publicación
2022-03-15Editor
The Royal Society of ChemistryCita bibliográfica
GALMÉS, Miquel À., et al. Computational design of an amidase by combining the best electrostatic features of two promiscuous hydrolases. Chemical Science, 2022, vol. 13, no 17, p. 4779-4787.Tipo de documento
info:eu-repo/semantics/articleVersión
info:eu-repo/semantics/publishedVersionPalabras clave / Materias
Resumen
While there has been emerging interest in designing new enzymes to solve practical challenges, computer-based options to redesign catalytically active proteins are rather limited. Here, a rational QM/MM molecular ... [+]
While there has been emerging interest in designing new enzymes to solve practical challenges, computer-based options to redesign catalytically active proteins are rather limited. Here, a rational QM/MM molecular dynamics strategy based on combining the best electrostatic properties of enzymes with activity in a common reaction is presented. The computational protocol has been applied to the re-design of the protein scaffold of an existing promiscuous esterase from Bacillus subtilis Bs2 to enhance its secondary amidase activity. After the alignment of Bs2 with a non-homologous amidase Candida antarctica lipase B (CALB) within rotation quaternions, a relevant spatial aspartate residue of the latter was transferred to the former as a means to favor the electrostatics of transition state formation, where a clear separation of charges takes place. Deep computational insights, however, revealed a significant conformational change caused by the amino acid replacement, provoking a shift in the pKa of the inserted aspartate and counteracting the anticipated catalytic effect. This prediction was experimentally confirmed with a 1.3-fold increase in activity. The good agreement between theoretical and experimental results, as well as the linear correlation between the electrostatic properties and the activation energy barriers, suggest that the presented computational-based investigation can transform in an enzyme engineering approach. [-]
Publicado en
Chem. Sci., 2022,13Entidad financiadora
Ministerio de Ciencia, Innovación y Universidades (Spain) | Generalitat Valenciana | Universitat Jaume I | Cardiff University | BBSRC | Leverhulme Trust | Royal Society of Chemistry
Código del proyecto o subvención
PGC2018-094852-B-C21 | PID2019-107098RJ-I00 | AICO/2019/195 | SEJI/2020/007 | UJI-A2019-04 | UJI-B2020-03 | BB/T015799/1 | RPG-2017-195 | RG170187 | RYC2020-030596-I | PREDOC/2017/23
Derechos de acceso
© 2022 The Author(s). Published by the Royal Society of Chemistry
info:eu-repo/semantics/openAccess
info:eu-repo/semantics/openAccess
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