title: Rational nanoconjugation improves biocatalytic performance of enzymes: aldol addition catalyzed by immobilized rhamnulose-1-phosphate aldolase.
abstract: Gold nanoparticles (AuNPs) are attractive materials for the immobilization of enzymes due to several advantages such as high enzyme loading, absence of internal diffusion limitations, and Brownian motion in solution, compared to the conventional immobilization onto porous macroscopic supports. The affinity of AuNPs to different groups present at the protein surface enables direct enzyme binding to the nanoparticle without the need of any coupling agent. Enzyme activity and stability appear to be improved when the biocatalyst is immobilized onto AuNPs. Rhamnulose-1-phosphate aldolase (RhuA) was selected as model enzyme for the immobilization onto AuNPs. The enzyme loading was characterized by four different techniques: surface plasmon resonance (SPR) shift and intensity, dynamic light scattering (DLS), and transmission electron microscopy (TEM). AuNPs-RhuA complexes were further applied as biocatalyst of the aldol addition reaction between dihydroxyacetone phosphate (DHAP) and (S)-Cbz-alaninal during two reaction cycles. In these conditions, an improved reaction yield and selectivity, together with a fourfold activity enhancement were observed, as compared to soluble RhuA.
authors.affiliation: Departament d'Enginyeria Química, Universitat Autònoma de Barcelona, Unitat de Biocatàlisi Aplicada associada al IQAC (UAB-CSIC), Barcelona, Spain.
authors.forename: Inés
authors.initials: I
authors.lastname: Ardao
authors.forename: Joan
authors.initials: J
authors.lastname: Comenge
authors.forename: M Dolors
authors.initials: MD
authors.lastname: Benaiges
authors.forename: Gregorio
authors.initials: G
authors.lastname: Álvaro
authors.forename: Víctor F
authors.initials: VF
authors.lastname: Puntes
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives / Q000378:metabolism; D000446:Aldehyde-Lyases / Q000737:chemistry* / Q000378:metabolism*; D000447:Aldehydes / Q000737:chemistry*; D055162:Biocatalysis; D004099:Dihydroxyacetone Phosphate / Q000378:metabolism; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D058726:Nanoconjugates / Q000737:chemistry*; D012210:Rhamnose / Q000031:analogs & derivatives / Q000378:metabolism
chemical_list: D000447:Aldehydes; D004800:Enzymes, Immobilized; C086275:N-carbobenzyloxyalanine; D058726:Nanoconjugates; C010419:rhamnulose 1-phosphate; D004099:Dihydroxyacetone Phosphate; D006046:Gold; C116609:3-hydroxybutanal; D000446:Aldehyde-Lyases; C022526:rhamnulose-1-phosphate adolase; D000409:Alanine; D012210:Rhamnose