dc.contributor.author
Land, Henrik
dc.contributor.author
Ceccaldi, Pierre
dc.contributor.author
Mészáros, Lívia S.
dc.contributor.author
Lorenzi, Marco
dc.contributor.author
Redman, Holly J.
dc.contributor.author
Senger, Moritz
dc.contributor.author
Stripp, Sven T.
dc.contributor.author
Berggren, Gustav
dc.date.accessioned
2019-09-24T05:34:26Z
dc.date.available
2019-09-24T05:34:26Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/25616
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-25382
dc.description.abstract
A new screening method for [FeFe]-hydrogenases is described, circumventing the need for specialized expression conditions as well as protein purification for initial characterization. [FeFe]-hydrogenases catalyze the formation and oxidation of molecular hydrogen at rates exceeding 103 s-1, making them highly promising for biotechnological applications. However, the discovery of novel [FeFe]-hydrogenases is slow due to their oxygen sensitivity and dependency on a structurally unique cofactor, complicating protein expression and purification. Consequently, only a very limited number have been characterized, hampering their implementation. With the purpose of increasing the throughput of [FeFe]-hydrogenase discovery, we have developed a screening method that allows for rapid identification of novel [FeFe]-hydrogenases as well as their characterization with regards to activity (activity assays and protein film electrochemistry) and spectroscopic properties (electron paramagnetic resonance and Fourier transform infrared spectroscopy). The method is based on in vivoartificial maturation of [FeFe]-hydrogenases in Escherichia coli and all procedures are performed on either whole cells or non-purified cell lysates, thereby circumventing extensive protein purification. The screening was applied on eight putative [FeFe]-hydrogenases originating from different structural sub-classes and resulted in the discovery of two new active [FeFe]-hydrogenases. The [FeFe]-hydrogenase from Solobacterium moorei shows high H2-gas production activity, while the enzyme from Thermoanaerobacter mathranii represents a hitherto uncharacterized [FeFe]-hydrogenase sub-class. This latter enzyme is a putative sensory hydrogenase and our in vivo spectroscopy study reveals distinct differences compared to the well established H2 producing HydA1 hydrogenase from Chlamydomonas reinhardtii.
en
dc.format.extent
9 S. (Manuskriptversion)
dc.rights.uri
https://creativecommons.org/licenses/by/4.0/
dc.subject
Novel [FeFe]-Hydrogenases
en
dc.subject
Biocatalytic H2-production
en
dc.subject.ddc
500 Naturwissenschaften und Mathematik::540 Chemie::540 Chemie und zugeordnete Wissenschaften
dc.title
Discovery of Novel [FeFe]-Hydrogenases for Biocatalytic H2-production
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation.articlenumber
xx
dcterms.bibliographicCitation.doi
10.1039/C9SC03717A
dcterms.bibliographicCitation.journaltitle
Chemical Science
dcterms.bibliographicCitation.number
xx
dcterms.bibliographicCitation.volume
2019
refubium.affiliation
Physik
refubium.note.author
Bei der PDF-Datei handelt es sich um eine Manuskriptversion.
refubium.resourceType.isindependentpub
no
dcterms.accessRights.openaire
open access
dcterms.isPartOf.issn
2041-6520
dcterms.isPartOf.eissn
2041-6539