Fachgebiet Physikalische Chemie / Biophysikalische Chemie

19 Items

Recent Submissions
Nuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O-2-tolerant NAD(+)-reducing [NiFe] hydrogenase

Lauterbach, Lars ; Wang, Hongxin ; Horch, Marius ; Gee, Leland B. ; Yoda, Yoshitaka ; Tanaka, Yoshihito ; Zebger, Ingo ; Lenz, Oliver ; Cramer, Stephen P. (2015)

Hydrogenases are complex metalloenzymes that catalyze the reversible splitting of molecular hydrogen into protons and electrons essentially without overpotential. The NAD(+)-reducing soluble hydrogenase (SH) from Ralstonia eutropha is capable of H-2 conversion even in the presence of usually toxic dioxygen. The molecular details of the underlying reactions are largely unknown, mainly because of...

Surface enhanced vibrational spectroscopic evidence for an alternative DNA-independent redox activation of endonuclease III

Moe, Elin ; Sezer, Murat ; Hildebrandt, Peter ; Todorovic, Smilja (2015)

Surface enhanced vibrational spectro-electrochemistry of endonuclease III provides direct evidence that the [4Fe-4S] cluster is responsible for the enzyme redox activity, and that this process is not exclusively DNA-mediated, as currently proposed. We report the first surface enhanced resonance Raman spectrum of a [4Fe-4S](2+) cluster containing enzyme.

Microporous polymer network films covalently bound to gold electrodes

Becker, Daniel ; Heidary, Nina ; Horch, Marius ; Gernert, Ulrich ; Zebger, Ingo ; Schmidt, Johannes ; Fischer, Anna ; Thomas, Arne (2015)

Covalent attachment of a microporous polymer network (MPN) on a gold surface is presented. A functional bromophenyl-based self-assembled monolayer (SAM) formed on the gold surface acts as co-monomer in the polymerisation of the MPN yielding homogeneous and robust coatings. Covalent binding of the films to the electrode is confirmed by SEIRAS measurements.

The influence of poly(ethylene glycol) on the micelle formation of alkyl maltosides used in membrane protein crystallization

Müh, Frank ; DiFiore, Dörte ; Zouni, Athina (2015)

With the aim of better understanding the phase behavior of alkyl maltosides (n-alkyl-beta-D-maltosides, C(n)G(2)) under the conditions of membrane protein crystallization, we studied the influence of poly(ethylene glycol) (PEG) 2000, a commonly used precipitating agent, on the critical micelle concentration (CMC) of the alkyl maltosides by systematic variation of the number n of carbon atoms in...

Cofactor composition and function of a H2-sensing regulatory hydrogenase as revealed by Mossbauer and EPR spectroscopy

Roncaroli, Federico ; Bill, Eckhard ; Friedrich, Bärbel ; Lenz, Oliver ; Lubitz, Wolfgang ; Pandelia, Maria-Eirini (2015)

The regulatory hydrogenase (RH) from Ralstonia eutropha H16 acts as a sensor for the detection of environmental H-2 and regulates gene expression related to hydrogenase-mediated cellular metabolism. In marked contrast to prototypical energy-converting [NiFe] hydrogenases, the RH is apparently insensitive to inhibition by O-2 and CO. While the physiological function of regulatory hydrogenases is...

Concepts in bio-molecular spectroscopy: vibrational case studies on metalloenzymes

Horch, Marius ; Hildebrandt, Peter ; Zebger, Ingo (2015)

Spectroscopic techniques play a major role in the elucidation of structure-function relationships of biological macromolecules. Here we describe an integrated approach for bio-molecular spectroscopy that takes into account the special characteristics of such compounds. The underlying fundamental concepts will be exemplarily illustrated by means of selected case studies on biocatalysts, namely h...

Calculating average surface enhancement factors of randomly nanostructured electrodes by a combination of SERS and impedance spectroscopy

Kozuch, Jacek ; Petrusch, N. ; Gkogkou, Dimitra ; Gernert, Ulrich ; Weidinger, Inez M. (2015)

Polyhedron Ag nanostructures were created on top of a polished Au electrode via step-wise electrodeposition and tested as substrates for SERS spectroscopy. Average Raman enhancement factors were derived by combining SERS measurements with electrochemical impedance spectroscopy (EIS), which is able to determine the electroactive surface area of a randomly nanostructured surface. Depending on the...

H-2-driven biotransformation of n-octane to 1-octanol by a recombinant Pseudomonas putida strain co-synthesizing an O-2-tolerant hydrogenase and a P450 monooxygenase

Lonsdale, Thomas H. ; Lauterbach, Lars ; Malca, Sumire Honda ; Nestl, Bettina M. ; Hauer, Bernhard ; Lenz, Oliver (2015)

An in vivo biotransformation system is presented that affords the hydroxylation of n-octane to 1-octanol on the basis of NADH-dependent CYP153A monooxygenase and NAD(+)-reducing hydrogenase heterologously synthesized in a bacterial host. The hydrogenase sustains H-2-driven NADH cofactor regeneration even in the presence of O-2, the co-substrate of monooxygenase.

2nd coordination sphere controlled electron transfer of iron hangman complexes on electrodes probed by surface enhanced vibrational spectroscopy

Ly, Hoang Khoa ; Wrzolek, Pierre ; Heidary, Nina ; Götz, R. ; Horch, Marius ; Kozuch, Jacek ; Schwalbe, Matthias ; Weidinger, Inez M. (2015)

Iron hangman complexes exhibit improved catalytic properties regarding O-2 and H2O2 reduction, which are attributed to the presence of a proton donating group in defined vicinity of the catalytic metal centre. Surface enhanced resonance Raman (SERR) and IR (SEIRA) spectro-electrochemistry has been applied concomitantly for the first time to analyse such iron hangman porphyrin complexes attached...

Reactivation from the Ni-B state in [NiFe] hydrogenase of Ralstonia eutropha is controlled by reduction of the superoxidised proximal cluster

Radu, Valentin ; Frielingsdorf, Stefan ; Lenz, Oliver ; Jeuken, Lars J. C. (2016)

The tolerance towards oxic conditions of O-2-tolerant [NiFe] hydrogenases has been attributed to an unusual [4Fe-3S] cluster that lies proximal to the [NiFe] active site. Upon exposure to oxygen, this cluster converts to a superoxidised (5+) state, which is believed to secure the formation of the so-called Ni-B state that is rapidly reactivated under reducing conditions. Here, the reductive rea...

Characterization of the [3Fe-4S](0/1+) cluster from the D14C variant of Pyrococcus furiosus ferredoxin via combined NRVS and DFT analyses

Lauterbach, Lars ; Gee, Leland B. ; Pelmenschikov, Vladimir ; Jenney, Francis E., Jr. ; Kamali, Saeed ; Yoda, Yoshitaka ; Adams, Michael W. W. ; Cramer, Stephen P. (2016)

The D14C variant of Pyrococcus furiosus ferredoxin provides an extraordinary framework to investigate a [3Fe-4S] cluster at two oxidation levels and compare the results to its physiologic [4Fe-4S] counterpart in the very same protein. Our spectroscopic and computational study reveals vibrational property changes related to the electronic and structural aspects of both Fe-S clusters.

When the inhibitor tells more than the substrate: the cyanide-bound state of a carbon monoxide dehydrogenase

Ciaccafava, Alexandre ; Tombolelli, Daria ; Domnik, Lilith ; Fesseler, Jochen ; Jeoung, Jae-Hun ; Dobbek, Holger ; Mroginski, Maria Andrea ; Zebger, Ingo ; Hildebrandt, Peter (2016)

Carbon monoxide dehydrogenase (CODH) is a key enzyme for reversible CO interconversion. To elucidate structural and mechanistic details of CO binding at the CODH active site (C-cluster), cyanide is frequently used as an iso-electronic substitute and inhibitor. However, previous studies revealed conflicting results on the structure of the cyanide-bound complex and the mechanism of cyanide-inhibi...

Changing the chemical and physical properties of high valent heterobimetallic bis-(mu-oxido) Cu-Ni complexes by ligand effects

Kafentzi, Maria-Chrysanthi ; Orio, Maylis ; Réglier, Marius ; Yao, Shenglai ; Kuhlmann, Uwe ; Hildebrandt, Peter ; Drieß, Matthias ; Simaan, A. Jalila ; Ray, Kallol (2016)

Two new heterobimetallic [LNiO2Cu(RPY2)](+) (RPY2 = N-substituted bis 2-pyridyl(ethylamine) ligands with R = indane, 3a or R = Me, 3b) complexes have been spectroscopically trapped at low temperatures. They were prepared by reacting the mononuclear side-on LNiII superoxo precursor bearing a beta-diketiminate ligand (L = [HC-(CMeNC6H3(iPr)(2))(2)]) with the Cu(I) complexes. In contrast to the ox...

Vibrational spectroscopy reveals the initial steps of biological hydrogen evolution

Katz, Sagie ; Noth, Jens ; Horch, Marius ; Shafaat, Hannah ; Happe, Thomas ; Hildebrandt, Peter ; Zebger, Ingo (2016)

[FeFe] hydrogenases are biocatalytic model systems for the exploitation and investigation of catalytic hydrogen evolution. Here, we used vibrational spectroscopic techniques to characterize, in detail, redox transformations of the [FeFe] and [4Fe4S] sub-sites of the catalytic centre (H-cluster) in a monomeric [FeFe] hydrogenase. Through the application of low-temperature resonance Raman spectro...

Domain motions and electron transfer dynamics in 2Fe-superoxide reductase

Horch, Marius ; Utesch, Tillmann ; Hildebrandt, Peter ; Mroginski, Maria Andrea ; Zebger, Ingo (2016)

Superoxide reductases are non-heme iron enzymes that represent valuable model systems for the reductive detoxification of reactive oxygen species. In the present study, we applied different theoretical methods to study the structural dynamics of a prototypical 2Fe-superoxide reductase and its influence on electron transfer towards the active site. Using normal mode and essential dynamics analys...

Redox processes of heme proteins and metalloporphyrins studied by vibrational spectroelectrochemistry

Kielb, Patrycja (2017)

Heme proteins belong to the most abundant and widely used metallo-proteins throughout the biosphere. Their biocatalytic, electron transporting and oxygen-binding functions are ubiquitous in many life-sustaining reactions. The structure of a heme cofactor and the nature of the heme pocket are determinants of the protein’s role and its involvement in metabolic pathways. Immobilized on an electrod...

NAD(H)-coupled hydrogen cycling - structure-function relationships of bidirectional [NiFe] hydrogenases

Horch, Marius ; Lauterbach, Lars ; Lenz, Oliver ; Hildebrandt, Peter ; Zebger, Ingo (2012)

Hydrogenases catalyze the activation or production of molecular hydrogen. Due to their potential importance for future biotechnological applications, these enzymes have been in the focus of intense research for the past decades. Bidirectional [NiFe] hydrogenases are of particular interest as they couple the reversible cleavage of hydrogen to the redox conversion of NAD(H). In this account, we r...

Enzyme-modified particles for selective biocatalytic hydrogenation by hydrogen-driven NADH recycling

Reeve, Holly A. ; Lauterbach, Lars ; Lenz, Oliver ; Vincent, Kylie A. (2015)

We describe a new approach to selective H-2-driven hydrogenation that exploits a sequence of enzymes immobilised on carbon particles. We used a catalyst system that comprised alcohol dehydrogenase, hydrogenase and an NAD(+) reductase on carbon black to demonstrate a greater than 98% conversion of acetophenone to phenylethanol. Oxidation of H-2 by the hydrogenase provides electrons through the c...

Multilayered lipid membrane stacks for biocatalysis using membrane enzymes

Heath, George R. ; Li, Mengqiu ; Rong, Honling ; Radu, Valentin ; Frielingsdorf, Stefan ; Lenz, Oliver ; Butt, Julea N. ; Jeuken, Lars J. C. (2017)

Multilayered or stacked lipid membranes are a common principle in biology and have various functional advantages compared to single-lipid membranes, such as their ability to spatially organize processes, compartmentalize molecules, and greatly increase surface area and hence membrane protein concentration. Here, a supramolecular assembly of a multilayered lipid membrane system is reported in wh...