An in vitro reconstitution system to monitor iron transfer to the active site during the maturation of [NiFe]-hydrogenase

Evidence the Isc iron–sulfur cluster biogenesis machinery is the source of iron for [NiFe]-cofactor biosynthesis in Escherichia coli

“…Using native mass spectrometry (native MS), it was recently shown for a HypC paralogue in E. coli , termed HybG, that it interacts specifically with the apo-catalytic subunit of the H 2 -oxidising Hyd-2 enzyme 16 . This corroborates an earlier proposal 15 that the HypC family of proteins (which includes HybG) functions to transfer the Fe(CN) 2 CO group from the HypCD (or HybG-HypD) complex to its cognate apo-catalytic subunit 16 , 17 ; for example, in E. coli , HypC delivers the Fe(CN) 2 CO group specifically to apo-HycE, the precursor of the Hyd forming the H 2 -producing formate hydrogenlyase (FHL) complex 1 . Once this moiety has been introduced into the empty active site cavity, two other Hyp proteins (HypA and HypB) deliver the nickel ion to complete cofactor assembly and insertion 1 – 3 .…”

Evidence that the Isc Iron-Sulfur Cluster Biogenesis Machinery Delivers Iron for [NiFe]-Cofactor Biosynthesis inEscherichia coli

“…Using native mass spectrometry (native MS), it was recently shown for a HypC paralogue in E. coli , termed HybG, that it interacts specifically with the apo-catalytic subunit of the H 2 -oxidising Hyd-2 enzyme [15]. This corroborates an earlier proposal [14] that the HypC family of proteins (which includes HybG) functions to transfer the Fe(CN) 2 CO group from the HypCD (or HybG-HypD) complex to its cognate apo-catalytic subunit [15, 16]; for example, in E. coli , HypC delivers the Fe(CN) 2 CO group specifically to apo-HycE, the precursor of the Hyd forming the H 2 -producing formate hydrogenlyase (FHL) complex [1]. Once this moiety has been introduced into the empty active site cavity, two other Hyp proteins (HypA and HypB) deliver the nickel ion to complete cofactor assembly and insertion [1–3].…”

“…The copyright holder for this this version posted November 17, 2023. ; https://doi.org/10.1101/2023.11.17.567542 doi: bioRxiv preprint 4 (which includes HybG) functions to transfer the Fe(CN)2CO group from the HypCD (or HybG-HypD) complex to its cognate apo-catalytic subunit [15,16]; for example, in E. coli, HypC delivers the Fe(CN)2CO group specifically to apo-HycE, the precursor of the Hyd forming the H2-producing formate hydrogenlyase (FHL) complex [1]. Once this moiety has been introduced into the empty active site cavity, two other Hyp proteins (HypA and HypB) deliver the nickel ion to complete cofactor assembly and insertion [1][2][3].…”

Synthetic engineering of a new biocatalyst encapsulating [NiFe]-hydrogenases for enhanced hydrogen production

“…35–39 HyaC is an integral membrane cytochrome b subunit that is responsible for anchoring Ec Hyd-1 to the cytoplasmic membrane. 40 HyaD, HyaE and HyaF were assumed to be involved in hydrogenase biosynthesis and maturation, 3,37,41,42 but their exact roles are still poorly understood. Previous studies have shown that HyaAB-only assemblies exhibit H 2 -producing activities and there was no significant difference in H 2 production between E. coli cells expressing hyaABCDEF and hyaAB only.…”