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Retuning the catalytic bias and overpotential of a [NiFe]-hydrogenase via a single amino acid exchange at the electron entry/exit site

Abstract:
The redox chemistry of the electron entry/exit site in Escherichia coli hydrogenase-1 is shown to play a vital role in tuning biocatalysis. Inspired by nature, we generate a HyaA-R193L variant to disrupt a proposed Arg-His cation-π interaction in the secondary coordination sphere of the outermost, "distal", iron-sulfur cluster. This rewires the enzyme, enhancing the relative rate of H2 production and the thermodynamic efficiency of H2 oxidation catalysis. On the basis of Fourier transformed alternating current voltammetry measurements, we relate these changes in catalysis to a shift in the distal [Fe4S4]2+/1+ redox potential, a previously experimentally inaccessible parameter. Thus, metalloenzyme chemistry is shown to be tuned by the second coordination sphere of an electron transfer site distant from the catalytic center.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1021/jacs.7b03611

Authors


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Institution:
University of Oxford
Division:
MPLS
Department:
Computer Science
Role:
Author


More from this funder
Funding agency for:
Gavaghan, D
Bond, A
Parkin, A
Grant:
EP/I017909/1
DP170101535
DP170101535
More from this funder
Funding agency for:
Walton, J
Parkin, A
Grant:
EP/M506394/1
DP170101535
More from this funder
Funding agency for:
Walton, J
Gavaghan, D
Roessler, M
Grant:
EP/M506394/1
EP/I017909/1
EP/M024393/1
More from this funder
Funding agency for:
Flanagan, L
Grant:
WT095024MA
More from this funder
Funding agency for:
Adamson, H
Bond, A
Parkin, A
Grant:
BB/F017316/1
DP170101535
DP170101535


Publisher:
American Chemical Society
Journal:
Journal of the American Chemical Society More from this journal
Volume:
139
Issue:
31
Pages:
10677-10686
Publication date:
2017-07-12
Acceptance date:
2017-07-11
DOI:
EISSN:
1520-5126
ISSN:
0002-7863
Pmid:
28697596


Language:
English
Keywords:
Pubs id:
pubs:708065
UUID:
uuid:aaa5c831-ee6f-424e-adb7-3abd92046afb
Local pid:
pubs:708065
Source identifiers:
708065
Deposit date:
2017-08-22

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