Bio-Electrosynthesis of Vectorially Imprinted Polymer Nanofilms for Cytochrome P450cam

Katharina J. Jetzschmann, Steffen Tank, Gyula Jágerszki, R. Gyurcsányi, Ulla Wollenberger, Frieder W. Scheller

Research output: Contribution to journalArticle

Abstract

A new approach for synthesizing a “vectorially” imprinted polymer (VIP) is presented for the microbial cytochrome P450cam enzyme. A surface attached binding motif of a natural reaction partner of the target protein, putidaredoxin (Pdx), is the anchor to the underlying transducer. The 15 amino acid peptide anchor, which stems from the largest continuous amino acid chain within the binding site of Pdx was modified: (i) internal cysteines were replaced by serines to prevent disulfide bond formation; (ii) 2 ethylene glycol units were attached to the N-terminus as a spacer region; and (iii) an N-terminal cysteine was added to allow the immobilization on the gold electrode surface. Immobilization on GCE was achieved via an N-(1-pyrenyl)maleimide (NPM) cross-linker. In this way oriented immobilization of P450cam was accomplished by binding it to a peptide-modified gold or glassy carbon electrode (GCE) prior to the electrosynthesis of a polymer nanofilm around the immobilized target. This VIP nanofilm enabled reversible oriented docking of P450cam as it is indicated by the catalytic oxygen reduction via direct electron transfer between the enzyme and the underlying electrode. Catalysis of oxygen reduction by P450cam bound to the VIP-modified GCE was used to measure rebinding to the VIP. The “mild” coupling of an oxidoreductase with the electrode may be appropriate for realizing electrode-driven substrate conversion by instable P450 enzymes without the need of NADPH co-factor.

Original languageEnglish
JournalChemElectroChem
DOIs
Publication statusPublished - Jan 1 2019

Fingerprint

Camphor 5-Monooxygenase
Cytochromes
Polymers
Proteins
Electrodes
Glassy carbon
Anchors
Gold
Peptides
Cysteine
Amino acids
Enzymes
Oxygen
Amino Acids
Ethylene Glycol
Binding sites
Ethylene glycol
NADP
Disulfides
Cytochrome P-450 Enzyme System

Keywords

  • cytochrome P450
  • direct electron transfer
  • electropolymerization
  • molecularly imprinted polymers
  • protein imprinting

ASJC Scopus subject areas

  • Catalysis
  • Electrochemistry

Cite this

Bio-Electrosynthesis of Vectorially Imprinted Polymer Nanofilms for Cytochrome P450cam. / Jetzschmann, Katharina J.; Tank, Steffen; Jágerszki, Gyula; Gyurcsányi, R.; Wollenberger, Ulla; Scheller, Frieder W.

In: ChemElectroChem, 01.01.2019.

Research output: Contribution to journalArticle

Jetzschmann, Katharina J. ; Tank, Steffen ; Jágerszki, Gyula ; Gyurcsányi, R. ; Wollenberger, Ulla ; Scheller, Frieder W. / Bio-Electrosynthesis of Vectorially Imprinted Polymer Nanofilms for Cytochrome P450cam. In: ChemElectroChem. 2019.
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