Phototrophic hydrogen production from a clostridial [FeFe] hydrogenase expressed in the heterocysts of the cyanobacterium Nostoc PCC 7120
- 325 Downloads
The conversion of solar energy into hydrogen represents a highly attractive strategy for the production of renewable energies. Photosynthetic microorganisms have the ability to produce H2 from sunlight but several obstacles must be overcome before obtaining a sustainable and efficient H2 production system. Cyanobacteria harbor [NiFe] hydrogenases required for the consumption of H2. As a result, their H2 production rates are low, which makes them not suitable for a high yield production. On the other hand, [FeFe] enzymes originating from anaerobic organisms such as Clostridium exhibit much higher H2 production activities, but their sensitivity to O2 inhibition impairs their use in photosynthetic organisms. To reach such a goal, it is therefore important to protect the hydrogenase from O2. The diazotrophic filamentous cyanobacteria protect their nitrogenases from O2 by differentiating micro-oxic cells called heterocysts. Producing [FeFe] hydrogenase in the heterocyst is an attractive strategy to take advantage of their potential in a photosynthetic microorganism. Here, we present a biological engineering approach for producing an active [FeFe] hydrogenase (HydA) from Clostridium acetobutylicum in the heterocysts of the filamentous cyanobacterium Nostoc PCC7120. To further decrease the O2 amount inside the heterocyst, the GlbN cyanoglobin from Nostoc commune was coproduced with HydA in the heterocyst. The engineered strain produced 400 μmol-H2 per mg Chlorophyll a, which represents 20-fold the amount produced by the wild type strain. This result is a clear demonstration that it is possible to associate oxygenic photosynthesis with H2 production by an O2-sensitive hydrogenase.
KeywordsClostridium acetobutylicum Cyanobacteria Cyanoglobin Heterocyst Hydrogen Nostoc
The authors thank Cheng-Cai Zhang for helpful discussions and Regine Lebrun from the “Plateforme Protéomique, FR3479 IMM” for the mass spectrometry analysis. We thank Jessica Blanc for revising the English manuscript. This research was supported by the “Agence Nationale pour la Recherche Scientifique” (ANR-13-BIME-0001).
Compliance with ethical standards
The authors declare that they have no competing interests.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Boughanemi S, Lyonnet J, Infossi P, Bauzan M, Kosta A, Lignon S, Giudici-Orticoni MT, Guiral M (2016) Microbial oxidative sulfur metabolism: biochemical evidence of the membrane-bound heterodisulfide reductase-like complex of the bacterium Aquifex aeolicus. FEMS Microbiol Lett 363(15). https://doi.org/10.1093/femsle/fnw156
- Chen CC, Lin CY, Chang JS (2001) Kinetics of hydrogen production with continuous anaerobic cultures utilizing sucrose as the limiting substrate. App Microbiol Biotechnol 57(1–2):56–64Google Scholar
- Gutekunst K, Chen X, Schreiber K, Kaspar U, Makam S, Appel J (2014) The bidirectional NiFe-hydrogenase in Synechocystis sp. PCC 6803 is reduced by flavodoxin and ferredoxin and is essential under mixotrophic, nitrate-limiting conditions. J Biol Chem 289(4):1930–1937. https://doi.org/10.1074/jbc.M113.526376 CrossRefPubMedGoogle Scholar
- Jeong JY, Yim HS, Ryu JY, Lee HS, Lee JH, Seen DS, Kang SG (2012) One-step sequence- and ligation-independent cloning as a rapid and versatile cloning method for functional genomics studies. Appl Environ Microbiol 78(15):5440–5443. https://doi.org/10.1128/AEM.00844-12 CrossRefPubMedPubMedCentralGoogle Scholar
- Kaneko T, Nakamura Y, Wolk CP, Kuritz T, Sasamoto S, Watanabe A, Iriguchi M, Ishikawa A, Kawashima K, Kimura T, Kishida Y, Kohara M, Matsumoto M, Matsuno A, Muraki A, Nakazaki N, Shimpo S, Sugimoto M, Takazawa M, Yamada M, Yasuda M, Tabata S (2001) Complete genomic sequence of the filamentous nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120. DNA Res 8(5):205–213 227-53CrossRefPubMedGoogle Scholar
- Nolling J, Breton G, Omelchenko MV, Makarova KS, Zeng Q, Gibson R, Lee HM, Dubois J, Qiu D, Hitti J, Wolf YI, Tatusov RL, Sabathe F, Doucette-Stamm L, Soucaille P, Daly MJ, Bennett GN, Koonin EV, Smith DR (2001) Genome sequence and comparative analysis of the solvent-producing bacterium Clostridium acetobutylicum. J Bacteriol 183(16):4823–4838. https://doi.org/10.1128/JB.183.16.4823-4838.2001 CrossRefPubMedPubMedCentralGoogle Scholar
- Ungerer JL, Pratte BS, Thiel T (2010) RNA Processing of Nitrogenase Transcripts in the Cyanobacterium Anabaena variabilis. J Bacteriol 192(13):3311-3320Google Scholar
- Zhao W, Ye Z, Zhao J (2007) RbrA, a cyanobacterial rubrerythrin, functions as a FNR-dependent peroxidase in heterocysts in protection of nitrogenase from damage by hydrogen peroxide in Anabaena sp. PCC 7120. Mol Microbiol 66(5):1219–1230. https://doi.org/10.1111/j.1365-2958.2007.05994.x CrossRefPubMedGoogle Scholar