TY  - JOUR
A1  - Jetzschmann, Katharina J.
A1  - Tank, Steffen
A1  - Jagerszki, Gyula
A1  - Gyurcsanyi, Robert E.
A1  - Wollenberger, Ulla
A1  - Scheller, Frieder W.
T1  - Bio-Electrosynthesis of Vectorially Imprinted Polymer Nanofilms for Cytochrome P450cam
JF  - ChemElectroChem
N2  - 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.
KW  - cytochrome P450
KW  - direct electron transfer
KW  - electropolymerization
KW  - molecularly imprinted polymers
KW  - protein imprinting
Y1  - 2019
U6  - https://doi.org/10.1002/celc.201801851
SN  - 2196-0216
VL  - 6
IS  - 6
SP  - 1818
EP  - 1823
PB  - Wiley-VCH
CY  - Weinheim
ER  -