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<p>Formate hydrogenlyases are metalloenzymes that can either produce molecular hydrogen gas or use H<sub>2</sub> to convert carbon dioxide to formic acid, thus potentially contributing doubly to a sustainable energy future. The structure of formate hydrogenlyase reveals a membrane-bound redox enzyme that shares a common ancestor with the mitochondrial complex I (NADH dehydrogenase). As such, formate hydrogenlyase falls into a category of so‐called ‘complex‐I‐like’ enzymes, that are found in bacteria, archaea and eukaryotic organelles. They share a common core structure of a membrane arm (most likely involved in proton or ion translocation) and a peripheral arm containing metal cofactors and involved in electron transfer. In this work, we clone a gene cluster from <i>Pectobacterium atrosepticum</i> encoding formate hydrogenlyase-2 (FHL-2). A bank of <i>Escherichia coli</i> host strains, themselves devoid of various combinations of native formate hydrogenlyase genes, are employed to characterise FHL-2. We demonstrate that <i>P. atrosepticum</i> FHL-2 is active in an <i>E. coli</i> host in that it can generate H<sub>2</sub> under fermentative growth conditions. Unlike native <i>E. coli</i> formate hydrogenlyase-1 (FHL-1), recombinant <i>P. atrosepticum</i> FHL-2 cannot perform the reverse reaction and generate formic acid from H<sub>2</sub> and CO<sub>2</sub>. By testing different combinations of genes by taking an in vivo cross-complementation approach we conclude that the extended membrane arm exhibited by FHL-2 is a major factor in controlling directionality of the enzyme.</p>

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Genetic dissection of formate hydrogenlyase-2 function: a metalloenzyme involved in bacterial hydrogen production

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  • 发表日期：

    2026年7月23日

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  • Thomas C.P. Reed,
  • Alexander J. Finney,
  • Magali Roger,
  • Frank Sargent

摘要

Formate hydrogenlyases are metalloenzymes that can either produce molecular hydrogen gas or use H2 to convert carbon dioxide to formic acid, thus potentially contributing doubly to a sustainable energy future. The structure of formate hydrogenlyase reveals a membrane-bound redox enzyme that shares a common ancestor with the mitochondrial complex I (NADH dehydrogenase). As such, formate hydrogenlyase falls into a category of so‐called ‘complex‐I‐like’ enzymes, that are found in bacteria, archaea and eukaryotic organelles. They share a common core structure of a membrane arm (most likely involved in proton or ion translocation) and a peripheral arm containing metal cofactors and involved in electron transfer. In this work, we clone a gene cluster from Pectobacterium atrosepticum encoding formate hydrogenlyase-2 (FHL-2). A bank of Escherichia coli host strains, themselves devoid of various combinations of native formate hydrogenlyase genes, are employed to characterise FHL-2. We demonstrate that P. atrosepticum FHL-2 is active in an E. coli host in that it can generate H2 under fermentative growth conditions. Unlike native E. coli formate hydrogenlyase-1 (FHL-1), recombinant P. atrosepticum FHL-2 cannot perform the reverse reaction and generate formic acid from H2 and CO2. By testing different combinations of genes by taking an in vivo cross-complementation approach we conclude that the extended membrane arm exhibited by FHL-2 is a major factor in controlling directionality of the enzyme.