Final Stages in the Biosynthesis of the [FeFe]-Hydrogenase Active Site

Xin Yu, Guodong Rao, R. David Britt, Thomas B. Rauchfuss

Research output: Contribution to journalArticlepeer-review


The paper aims to elucidate the final stages in the biosynthesis of the [2Fe]H active site of the [FeFe]-hydrogenases.  The recently hypothesized intermediate [Fe2(SCH2NH2)2(CN)2(CO)4]2- ([1]2-) was prepared by a multistep route from [Fe2(S2)(CN)(CO)5]-. The following synthetic intermediates were characterized in order: [Fe2(SCH2NHFmoc)2(CNBEt3)(CO)5]-, [Fe2(SCH2NHFmoc)2(CN)-(CO)5]-, and [Fe2(SCH2NHFmoc)2(CN)2(CO)4]2-, where Fmoc is  fluorenylmethoxycarbonyl). Derivatives of these anions include [K(18-crown-6)]+, PPh4+ and PPN+ salts as well as the 13CD2-isotopologues. These Fe2 species exist as a mixture of two isomers attributed to diequatorial (ee) and axial-equatorial (ae) stereochemistry at sulfur.  In vitro experiments demonstrate that [1]2- maturates HydA1 in the presence of HydF and a cocktail of reagents. HydA1 can also be maturated using a highly simplified cocktail, omitting HydF and other proteins. This result is consistent with HydA1 participating in the maturation process and refines the roles of HydF.
Original languageEnglish (US)
Article numbere202404044
JournalAngewandte Chemie International Edition
Issue number22
Early online dateMar 29 2024
StatePublished - May 27 2024


  • biosynthesis
  • hydrogenase
  • iron
  • electron-nuclear double resonance (ENDOR) spectroscopy
  • cofactor

ASJC Scopus subject areas

  • General Chemistry
  • Catalysis


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