Journal
NANOSCALE
Volume 4, Issue 2, Pages 659-666Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c1nr11443c
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Funding
- AOARD [FA2386-10-1-4065]
- AFOSR [FA9550-08-1-0124]
- DARPA
- DoD
- EPSRC [EP/F042590/1]
- Ministry of Social Justice and Empowerment, Government of India
- BBSRC [BB/I008055/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/I008055/1] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [EP/F042590/1] Funding Source: researchfish
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The development of a novel method for functionalizing nanopatterned surfaces with catalytically active proteins is reported. This method involves using dip-pen nanolithography (DPN) and polymer pen lithography (PPL) to generate nanoscale patterns of coenzyme A, followed by a phosphopantetheinyl transferase-mediated coupling between coenzyme A and proteins fused to the ybbR-tag. By exploiting the ability to generate protein features over large areas afforded by DPN and PPL, it was now possible to measure protein activity directly on these surfaces. It was found that proteins immobilized on the nanoscale features not only display higher activity per area with decreasing feature size, but are also robust and can be used for repeated catalytic cycles. The immobilization method is applicable to a variety of proteins and gives rise to superior activity compared to proteins attached in random orientations on the surface.
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