Site-selective immobilisation of functional enzymes on to polystyrene nanoparticles.
The immobilisation of proteins on to nanoparticles has a number of applications ranging from biocatalysis through to cellular delivery of biopharmaceuticals. Here we describe a phosphopantetheinyl transferase (Sfp)-catalysed method for immobilising proteins bearing a small 12-mer "ybbR" tag on to nanoparticles functionalised with coenzyme A. The Sfp-catalysed immobilisation of proteins on to nanoparticles is a highly efficient, single step reaction that proceeds under mild conditions and results in a homogeneous population of proteins that are covalently and site-specifically attached to the surface of the nanoparticles. Several enzymes of interest for biocatalysis, including an arylmalonate decarboxylase (AMDase) and a glutamate racemase (GluR), were immobilised on to nanoparticles using this approach. These enzymes retained their activity and showed high operational stability upon immobilisation.
School of Chemistry and Manchester Interdisciplinary Biocentre, The University of Manchester, 131 Princess Street, Manchester, UKM1 7DN.
Lu Shin
LS
Wong
Krzysztof
K
Okrasa
Jason
J
Micklefield
D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D002384:Catalysis; D004800:Enzymes, Immobilized / Q000008:administration & dosage / Q000378:metabolism*; D008958:Models, Molecular; D053758:Nanoparticles / Q000737:chemistry; D011137:Polystyrenes / Q000737:chemistry*; D017855:Transferases (Other Substituted Phosphate Groups) / Q000737:chemistry*
D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D011137:Polystyrenes; C082662:phosphopantetheinyl transferase; D017855:Transferases (Other Substituted Phosphate Groups)