Improved oxygen tolerance of the Synechocystis sp. PCC 6803 bidirectional hydrogenase by site-directed mutagenesis of putative residues of the gas diffusion channel

Cano, Mélissa; Volbeda, Anne; Guédeney, Geneviève; Aubert-Jousset, Emeline; Richaud, Pierre; Peltier, Gilles; Cournac, Laurent

doi:10.1016/j.ijhydene.2014.08.030

Cited by 17 publications

(14 citation statements)

References 49 publications

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“…However, group 5 [NiFe]-hydrogenases have found a way to overcome this: a range of in vitro and in vivo studies indicate that they are completely insensitive to O 2 (16,23,29 (40,41). The crystal structure of AH reveals that the gas diffusion channel is significantly narrower than that of the group 1 [NiFe]-hydrogenase, which may contribute to differences in O 2 susceptibility (30).…”

Section: Properties Of Group 5 [Nife]-hydrogenasesmentioning

confidence: 99%

Atmospheric Hydrogen Scavenging: from Enzymes to Ecosystems

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Constant

Hards

et al. 2015

Appl Environ Microbiol

100

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fWe have known for 40 years that soils can consume the trace amounts of molecular hydrogen (H 2 ) found in the Earth's atmosphere. This process is predicted to be the most significant term in the global hydrogen cycle. However, the organisms and enzymes responsible for this process were only recently identified. Pure culture experiments demonstrated that several species of Actinobacteria, including streptomycetes and mycobacteria, can couple the oxidation of atmospheric H 2 to the reduction of ambient O 2 . A combination of genetic, biochemical, and phenotypic studies suggest that these organisms primarily use this fuel source to sustain electron input into the respiratory chain during energy starvation. This process is mediated by a specialized enzyme, the group 5 [NiFe]-hydrogenase, which is unusual for its high affinity, oxygen insensitivity, and thermostability. Atmospheric hydrogen scavenging is a particularly dependable mode of energy generation, given both the ubiquity of the substrate and the stress tolerance of its catalyst. This minireview summarizes the recent progress in understanding how and why certain organisms scavenge atmospheric H 2 . In addition, it provides insight into the wider significance of hydrogen scavenging in global H 2 cycling and soil microbial ecology.

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Section: Properties Of Group 5 [Nife]-hydrogenasesmentioning

confidence: 99%

Atmospheric Hydrogen Scavenging: from Enzymes to Ecosystems

Greening

Constant

Hards

et al. 2015

Appl Environ Microbiol

100

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“…These genetic modifications were shown to be further associated with changes in the gas diffusion characteristics and they did affect the directionality of the enzyme (Abou Hamdan et al 2012). Recent reports have shown that a similar strategy could be developed on the hydrogenase of Synechocystis, resulting in the expression of an engineered hydrogenase exhibiting a sustained activity in the presence of O 2 with a bias toward production (Cano et al 2014). Other engineering strategies involving different targets can be considered and might even be required as part of a larger scale combination of unique redesigning in order to obtain an efficient O 2 -tolerant hydrogenase.…”

Section: Engineering O 2 -Tolerant [Nife]-hydrogenasesmentioning

confidence: 99%

Implementation of photobiological H2 production: the O2 sensitivity of hydrogenases

Ghirardi

2015

Photosynth Res

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The search for the ultimate carbon-free fuel has intensified in recent years, with a major focus on photoproduction of H2. Biological sources of H2 include oxygenic photosynthetic green algae and cyanobacteria, both of which contain hydrogenase enzymes. Although algal and cyanobacterial hydrogenases perform the same enzymatic reaction through metallo-clusters, their hydrogenases have evolved separately, are expressed differently (transcription of algal hydrogenases is anaerobically induced, while bacterial hydrogenases are constitutively expressed), and display different sensitivity to O2 inactivation. Among various physiological factors, the sensitivity of hydrogenases to O2 has been one of the major factors preventing implementation of biological systems for commercial production of renewable H2. This review addresses recent strategies aimed at engineering increased O2 tolerance into hydrogenases (as of now mainly unsuccessful), as well as towards the development of methods to bypass the O2 sensitivity of hydrogenases (successful but still yielding low solar conversion efficiencies). The author concludes with a description of current approaches from various laboratories to incorporate multiple genetic traits into either algae or cyanobacteria to jointly address limiting factors other than the hydrogenase O2 sensitivity and achieve more sustained H2 photoproduction activity.

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“…Therefore exchanges of V263Q, E340D and M362V were unlikely to affect the oxygen tolerance of the KHyd3 by altering the reductive potential of the Ni-Fe active center. Some studies deemed changing AARs of gas tunnel could alter [NiFe] hydrogenases' O 2sensitivity by blocking O 2 access to the Ni-Fe active center [15][16][17][18][19][20]. However, all effective ''gateway'' AARs found were next to the Ni-Fe active center.…”

Section: The Surface Aars Affect the Oxygen Tolerance Of Khyd3mentioning

confidence: 99%

“…One was that hydrogenases' O 2 -tolerance was mainly due to the effect of gas channels, which restricted O 2 access to active center via gas channels. Some studies demonstrated that certain hydrogenases were slowly inactivated by O 2 because the diffusion rate of O 2 to the active center via gas channels was lower [5,[13][14][15][16][17][18][19][20]. Some AARs which located at inner exit of gas channels next to the active-center were found.…”

Section: Introductionmentioning

confidence: 99%

“…Some AARs which located at inner exit of gas channels next to the active-center were found. They impacted hydrogenases' O 2 -tolerance by restricting O 2 access to active center via gas channels (diminished channel diameters) [15][16][17][18][19][20]. The other one was that hydrogenases' O 2 -tolerance mainly originated from the reductive removal of oxygenic species from oxygen-bound Ni-Fe active-center guided by a unique architecture of the electron relay.…”

Section: Introductionmentioning

confidence: 99%

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Improved O₂‐tolerance in variants of a H₂‐evolving [NiFe]‐hydrogenase from Klebsiella oxytoca HP1

Huang

Bai

et al. 2015

FEBS Letters

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a b s t r a c tIn this study, we investigated the mechanism of O 2 tolerance of Klebsiella oxytoca HP1 H 2 -evolving hydrogenase 3 (KHyd3) by mutational analysis and three-dimensional structure modeling. Results revealed that certain surface amino acid residues of KHyd3 large subunit, in particular those at the outer entrance of the gas channel, have a visible effect on its oxygen tolerance. Additionally, solution pH, immobilization and O 2 partial pressure also affect KHyd3 O 2 -tolerance to some extent. We propose that the extent of KHyd3 O 2 -tolerance is determined by a balance between the rate of O 2 access to the active center through gas channels and the deoxidation rate of the oxidized active center. Based on our findings, two higher O 2 -tolerant KHyd3 mutations G300E and G300M were developed.

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Improved oxygen tolerance of the Synechocystis sp. PCC 6803 bidirectional hydrogenase by site-directed mutagenesis of putative residues of the gas diffusion channel

Cited by 17 publications

References 49 publications

Atmospheric Hydrogen Scavenging: from Enzymes to Ecosystems

Atmospheric Hydrogen Scavenging: from Enzymes to Ecosystems

Implementation of photobiological H2 production: the O2 sensitivity of hydrogenases

Improved O₂‐tolerance in variants of a H₂‐evolving [NiFe]‐hydrogenase from Klebsiella oxytoca HP1

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Improved oxygen tolerance of the Synechocystis sp. PCC 6803 bidirectional hydrogenase by site-directed mutagenesis of putative residues of the gas diffusion channel

Cited by 17 publications

References 49 publications

Atmospheric Hydrogen Scavenging: from Enzymes to Ecosystems

Atmospheric Hydrogen Scavenging: from Enzymes to Ecosystems

Implementation of photobiological H2 production: the O2 sensitivity of hydrogenases

Improved O2‐tolerance in variants of a H2‐evolving [NiFe]‐hydrogenase from Klebsiella oxytoca HP1

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Improved O₂‐tolerance in variants of a H₂‐evolving [NiFe]‐hydrogenase from Klebsiella oxytoca HP1