TY  - JOUR
A1  - Yarman, Aysu
T1  - Electrosynthesized Molecularly Imprinted Polymer for Laccase Using the Inactivated Enzyme as the Target
JF  - Bulletin of the Korean chemical society
N2  - The first molecularly imprinted polymer (MIP) for the recognition of the copper-enzyme laccase was successfully prepared by electropolymerizing scopoletin in the presence of alkaline-inactivated enzyme. Laccase-MIP and the control polymer without laccase (nonimprinted polymer, NIP) were characterized by voltammetry using the redox marker ferricyanide. After electropolymerization, the signals for ferricyanide for both the MIP and the NIP were almost completely suppressed and increased after removal of the target from the polymer layer. Rebinding of both inactivated and active laccase decreased the ferricyanide peak currents to almost equal extent. The relative decrease of signal suppression approached saturation above 10 nM. Furthermore, the surface activity of rebound laccase toward the oxidation of catechol was investigated. The surface activity approached saturation above 10 nM, a value close to the value of the measurements with ferricyanide. Interaction of NIP with laccase brought about a six times smaller signal of catechol oxidation.
KW  - Molecularly imprinted polymers
KW  - Biomimetic sensors
KW  - Laccase
KW  - Electropolymerization
KW  - Scopoletin
Y1  - 2018
U6  - https://doi.org/10.1002/bkcs.11413
SN  - 1229-5949
VL  - 39
IS  - 4
SP  - 483
EP  - 488
PB  - Wiley-VCH
CY  - Weinheim
ER  - 
TY  - JOUR
A1  - Jetzschmann, Katharina J.
A1  - Yarman, Aysu
A1  - Rustam, L.
A1  - Kielb, P.
A1  - Urlacher, V. B.
A1  - Fischer, A.
A1  - Weidinger, I. M.
A1  - Wollenberger, Ulla
A1  - Scheller, Frieder W.
T1  - Molecular LEGO by domain-imprinting of cytochrome P450 BM3
JF  - Colloids and surfaces : an international journal devoted to fundamental and applied research on colloid and interfacial phenomena in relation to systems of biological origin ; B, Biointerfaces
N2  - Hypothesis: Electrosynthesis of the MIP nano-film after binding of the separated domains or holocytochrome BM3 via an engineered anchor should result in domain-specific cavities in the polymer layer. Experiments: Both the two domains and the holo P450 BM3 have been bound prior polymer deposition via a N-terminal engineered his6-anchor to the electrode surface. Each step of MIP preparation was characterized by cyclic voltammetry of the redox-marker ferricyanide. Rebinding after template removal was evaluated by quantifying the suppression of the diffusive permeability of the signal for ferricyanide and by the NADH-dependent reduction of cytochrome c by the reductase domain (BMR). Findings: The working hypothesis is verified by the discrimination of the two domains by the respective MIPs: The holoenzyme P450 BM3 was ca. 5.5 times more effectively recognized by the film imprinted with the oxidase domain (BMO) as compared to the BMR-MIP or the non-imprinted polymer (NIP). Obviously, a cavity is formed during the imprinting process around the hiss-tag-anchored BMR which cannot accommodate the broader BMO or the P450 BM3. The affinity of the MIP towards P450 BM3 is comparable with that to the monomer in solution. The hiss-tagged P450 BM3 binds (30 percent) stronger which shows the additive effect of the interaction with the MIP and the binding to the electrode.
KW  - Molecularly imprinted polymers
KW  - Protein imprinting
KW  - Electropolymerization
KW  - Cytochrome P450
Y1  - 2018
U6  - https://doi.org/10.1016/j.colsurfb.2018.01.047
SN  - 0927-7765
SN  - 1873-4367
VL  - 164
SP  - 240
EP  - 246
PB  - Elsevier
CY  - Amsterdam
ER  -