Congratulations Patri, Simone and all the team! Great paper!

Thank you to my amazing collaborators and the PhD student doing the work, Sam!
Please have a read, it is a really interesting paper!

An elegant engineering strategy to perform step-wise truncations of the iron-sulfur cluster domains in semi-synthetic [FeFe] hydrogenases to investigate in detail the influence on the catalytic properties of the enzyme and the inhibitors binding!

1️⃣ 𝗠𝗜𝗕𝗧𝗣-𝗕𝗕𝗦𝗥𝗖 - 𝟮𝟳𝘁𝗵 𝗡𝗼𝘃𝗲𝗺𝗯𝗲𝗿 𝟮𝟬𝟮𝟱 𝗱𝗲𝗮𝗱𝗹𝗶𝗻𝗲:
warwick.ac.uk/fac/cross_fa...

2️⃣ 𝗖𝗘𝗡𝗧𝗔-𝗡𝗘𝗥𝗖 - 𝟳𝘁𝗵 𝗝𝗮𝗻𝘂𝗮𝗿𝘆 𝟮𝟬𝟮𝟲 𝗱𝗲𝗮𝗱𝗹𝗶𝗻𝗲:
centa.ac.uk/studentship/...

Any questions or more information? Please email me, happy to help!

#metalloenzymes #biohybridcatalysts #sustainablechemistry #greenchemistry

Information about the project here: le.ac.uk/research/ins...

Information about the position and how to apply:
jobs.le.ac.uk/vacancies/11...

Deadline 7 Dec 2025!

For any enquires please send an email!

Ok, thanks. Solution potential of the activity assay ought to be pretty negative though, right? 100 mM sodium dithionite at pH 6.8. I’d guess ca. -600 mV vs SHE compared with an eq potential of H2 of ca. -400. So you could have 200 mV of overpotential…

I’m also interested in a coordination chemist’s view of the 7 coordinate Fe intermediate. Is there likely to be a low barrier to its formation?

Hi Sven. Interesting paper. You said “no significant extra driving force”…You mean no significant overpotential? What is the overpotential and how did you measure it? Also, is it right that the turnover frequency is less than 1 per second? Any ideas how to make this better?

We probed both directions, and it really seems to be the F-domain that modulates the on/off dynamics. We didn't investigate DdH tho...

We have positions for pCNF SDM of CpI in a recent grant application... Very encouraging to see Kylie's data!

Presumably, only reduced ferredoxin binds to *oxidised* hydrogenase (during proton reduction)? Meanwhile oxidised ferredoxin should bind to reduced hydrogenase (during hydrogen oxidation)? I guess I need to read the paper…nice work! Any plans to engineer pCNF into CpI?

We believe that these protein-protein-interactions propell fast & repeated one-electron-transfer reactions, driving multi-electron redox catalysis with a one-electron mediator. In hydrogenase, and other oxidoreductases as well.
Find all the details in the paper:
pubs.rsc.org/en/content/a...

Uli's brilliant microscale thermophoresis experiments show that *only* reduced ferredoxin binds to hydrogenase, which facilitates assigning the secondary structural changes to redox changes and a shift from an "attractive" to a "repulsive" configuration.

Infrared difference spectra reveal that the reduction of ferredoxin is accompanied with larger secondary structural changes - clearly associated with the "F-domain" of [FeFe]-hydrogenase, as reference experiments with "minimal hydrogenase" CrHydA1 lack this spectral feature.
A surprise‼️

Interestingly, in the ferredoxin from C. pasteurianum, reduction alone does *not* affect the nitrile frequency. Only in complex with [FeFe]-hydrogenase CpI, redox changes affect the artificial amino acid...