Chapter 20 : Towards Hydrogenase Engineering for Hydrogen Production

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This chapter discusses hydrogenase structure-function relationship studies in which new properties of modified enzymes might serve as an inspiration source for rational optimization of hydrogenases for biotechnological processes. A key point is that these reactions that appear as competitors for biotechnological purposes are often essential for cell survival or development. In part, this explains the difficulty and the slow progress in biohydrogen research. Two research directions can be proposed to overcome this kind of limitation: improve the substrate specificity of hydrogenase or, more radically, redirect redox intermediates. The V74M-L122M hydrogenase oxidized by oxygen remained in the same redox state as the native enzyme oxidized anaerobically, as demonstrated by the predominance of an Ni-B EPR signal (while Ni-A is predominant in the oxygen-exposed native enzyme) and by the abundance of a hydroxyl-bridging ligand at the active site in the structure. It was shown that it is possible to improve dioxygen resistance of [NiFe] hydrogenases. The enzyme bias, substrate specificity, and oxygen resistance are the main domains in which some progress has already been made, opening the way towards future applications. But other issues, like heterologous expression of [NiFe] hydrogenases that would facilitate molecular research and organism engineering or deciphering the catalytic mechanism that would allow the development of biomimetic catalysts, are also the subjects of intensive research and will contribute to biohydrogen implementation.

Citation: Rousset M, Cournac L. 2008. Towards Hydrogenase Engineering for Hydrogen Production, p 249-257. In Wall J, Harwood C, Demain A (ed), Bioenergy. ASM Press, Washington, DC. doi: 10.1128/9781555815547.ch20
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Image of Figure 1.
Figure 1.

[Fe] hydrogenase from ( ). Protein Data Bank ID 2b0j. Three-dimensional structure of the apoenzyme is shown.

Citation: Rousset M, Cournac L. 2008. Towards Hydrogenase Engineering for Hydrogen Production, p 249-257. In Wall J, Harwood C, Demain A (ed), Bioenergy. ASM Press, Washington, DC. doi: 10.1128/9781555815547.ch20
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Image of Figure 2.
Figure 2.

(Top row and bottom row, left and center) Photographs, taken through the optic microscope, of the bacterial cells presented in this chapter. (formerly ) is from Wiki microbe; is from Evolution Ecology and Biodiversity Lab Manual online from the University of Winnipeg; is from the Metalbioreduction web page; is from Viet Sciences; is from Wellesley College. (Bottom row, right) The photobioreactor is a bacteria reactor specially designed for the culture of photosynthetic organisms. The photobioreactor presented here contains growing cells from the Commissariat à l’Energie Atomique, the Institut de Biologie Environmentale et Biotechnologie, and the Laboratoire de Bioénergétique et Biotechnologie des Bactéries et Microalgues.

Citation: Rousset M, Cournac L. 2008. Towards Hydrogenase Engineering for Hydrogen Production, p 249-257. In Wall J, Harwood C, Demain A (ed), Bioenergy. ASM Press, Washington, DC. doi: 10.1128/9781555815547.ch20
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