Next Article in Journal
Deconstruction of the CYP153A6 Alkane Hydroxylase System: Limitations and Optimization of In Vitro Alkane Hydroxylation
Next Article in Special Issue
Construction of a Vitreoscilla Hemoglobin Promoter-Based Tunable Expression System for Corynebacterium glutamicum
Previous Article in Journal
Mg-Catalyzed OPPenauer Oxidation—Application to the Flow Synthesis of a Natural Pheromone
Previous Article in Special Issue
Efficient Conversion of Acetate to 3-Hydroxypropionic Acid by Engineered Escherichia coli
Article Menu
Issue 11 (November) cover image

Export Article

Open AccessArticle
Catalysts 2018, 8(11), 530;

Infrared Characterization of the Bidirectional Oxygen-Sensitive [NiFe]-Hydrogenase from E. coli

Department of Physics, Experimental Molecular Biophysics, Freie Universität Berlin, 14195 Berlin, Germany
Department of Physics, Genetic Biophysics, Freie Universität Berlin, 14195 Berlin, Germany
Author to whom correspondence should be addressed.
These authors contributed equally.
Received: 16 October 2018 / Revised: 31 October 2018 / Accepted: 6 November 2018 / Published: 8 November 2018
PDF [4809 KB, uploaded 8 November 2018]


[NiFe]-hydrogenases are gas-processing metalloenzymes that catalyze the conversion of dihydrogen (H2) to protons and electrons in a broad range of microorganisms. Within the framework of green chemistry, the molecular proceedings of biological hydrogen turnover inspired the design of novel catalytic compounds for H2 generation. The bidirectional “O2-sensitive” [NiFe]-hydrogenase from Escherichia coli HYD-2 has recently been crystallized; however, a systematic infrared characterization in the presence of natural reactants is not available yet. In this study, we analyze HYD-2 from E. coli by in situ attenuated total reflection Fourier-transform infrared spectroscopy (ATR FTIR) under quantitative gas control. We provide an experimental assignment of all catalytically relevant redox intermediates alongside the O2- and CO-inhibited cofactor species. Furthermore, the reactivity and mutual competition between H2, O2, and CO was probed in real time, which lays the foundation for a comparison with other enzymes, e.g., “O2-tolerant” [NiFe]-hydrogenases. Surprisingly, only Ni-B was observed in the presence of O2 with no indications for the “unready” Ni-A state. The presented work proves the capabilities of in situ ATR FTIR spectroscopy as an efficient and powerful technique for the analysis of biological macromolecules and enzymatic small molecule catalysis. View Full-Text
Keywords: redox enzymes; FTIR spectroscopy; small molecules redox enzymes; FTIR spectroscopy; small molecules

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material


Share & Cite This Article

MDPI and ACS Style

Senger, M.; Laun, K.; Soboh, B.; Stripp, S.T. Infrared Characterization of the Bidirectional Oxygen-Sensitive [NiFe]-Hydrogenase from E. coli. Catalysts 2018, 8, 530.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics



[Return to top]
Catalysts EISSN 2073-4344 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top