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Hydrogen photo-evolution upon S deprivation stepwise: an illustration of microalgal photosynthetic and metabolic flexibility and a step stone for future biotechnological methods of renewable H2 production

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Abstract

The metabolic flexibility of some photosynthetic microalgae enables them to survive periods of anaerobiosis in the light by developing a particular photofermentative metabolism. The latter entails compounds of the photosynthetic electron transfer chain and an oxygen-sensitive hydrogenase in order to reoxidize reducing equivalents and to generate ATP for maintaining basal metabolic function. This pathway results in the photo-evolution of hydrogen gas by the algae. A decade ago, Melis and coworkers managed to reproduce such a condition in a laboratory context by depletion of sulfur in the algal culture media, making the photo-evolution by the algae sustainable for several days (Melis et al. in Plant Physiol 122:127–136, 2000). This observation boosted research in algal H2 evolution. A feature, which due to its transient nature was long time considered as a curiosity of algal photosynthesis suddenly became a phenomenon with biotechnological potential. Although the Melis procedure has not been developed into a biotechnological process of renewable H2 generation so far, it has been a useful tool for studying microalgal metabolic and photosynthetic flexibility and a possible step stone for future H2 production procedures. Ten years later most of the critical steps and limitations of H2 production by this protocol have been studied from different angles particularly with the model organism Chlamydomonas reinhardtii, by introducing various changes in culture conditions and making use of mutants issued from different screens or by reverse genomic approaches. A synthesis of these observations with the most important conclusions driven from recent studies will be presented in this review.

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References

  • Abeles FB (1964) Cell-free hydrogenase from Chlamydomonas. Plant Physiol 39:169–176

    Article  CAS  PubMed  Google Scholar 

  • Antal TK, Krendeleva TE, Laurinavichene TV, Makarova VV, Ghirardi ML, Rubin AB, Tsygankov AA, Seibert M (2003) The dependence of algal H2 production on photosystem II and O2 consumption activities in sulfur-deprived Chlamydomonas reinhardtii cells. Biochim Biophys Acta 1607:153–160

    Article  CAS  PubMed  Google Scholar 

  • Antal TK, Volgusheva AA, Kukarskih GP, Krendeleva TE, Rubin AB (2009) Relationships between H2 photoproduction and different electron transport pathways in sulfur-deprived Chlamydomonas reinhardtii. Int J Hydrogen Energy 34:9087–9094

    Article  CAS  Google Scholar 

  • Bishop NI, Gaffron H (1963) On the interrelation of the mechanisms for oxygen and hydrogen evolution in adapted algae. In: Kok B, Jagendorf AT (eds) Photosynthetic mechanisms of green plants, Nat Acad Sci Publ 1145, pp 441–451

  • Cardol P, Gloire G, Havaux M, Remacle C, Matagne R, Franck F (2003) Photosynthesis and state transitions in mitochondrial mutants of Chlamydomonas reinhardtii affected in respiration. Plant Physiol 133:2010–2020

    Article  CAS  PubMed  Google Scholar 

  • Cardol P, Alric J, Girard-Bascou J, Franck F, Wollman F, Finazzi G (2009) Impaired respiration discloses the physiological significance of state transitions in Chlamydomonas. Proc Natl Acad Sci USA 106:15979–15984

    Article  CAS  PubMed  Google Scholar 

  • Chen H, Melis A (2004) Localization and function of SulP, a nuclear-encoded chloroplast sulfate permease in Chlamydomonas reinhardtii. Planta 220:198–210

    Article  CAS  PubMed  Google Scholar 

  • Chen H, Newton AJ, Melis A (2005) Role of SulP, a nuclear-encoded chloroplast sulfate permease, in sulfate transport and H2 evolution in Chlamydomonas reinhardtii. Photosynth Res 84:289–296

    Article  CAS  PubMed  Google Scholar 

  • Chochois V, Dauvillée D, Beyly A, Tolleter D, Cuiné S, Timpano H, Ball S, Cournac L, Peltier G (2009) Hydrogen production in Chlamydomonas: photosystem II-dependent and -independent pathways differ in their requirement for starch metabolism. Plant Physiol 151:631–640

    Article  CAS  PubMed  Google Scholar 

  • Cohen J, Kim K, Posewitz M, Ghirardi ML, Schulten K, Seibert M, King P (2005) Molecular dynamics and experimental investigation of H2 and O2 diffusion in [Fe]-hydrogenase. Biochem Soc Trans 33:80–82

    Article  CAS  PubMed  Google Scholar 

  • Cournac L, Mus F, Bernard L, Guedeney G, Vignais P, Peltier G (2002) Limiting steps of hydrogen production in Chlamydomonas reinhardtii and Synechocystis PCC 6803 as analysed by light-induced gas exchange transients. Int J Hydrogen Energy 27:1229–1237

    Article  CAS  Google Scholar 

  • Davies JP, Yildiz FH, Grossman AR (1999) Sac3, an Snf1-like serine/threonine kinase that positively and negatively regulates the responses of Chlamydomonas to sulfur limitation. Plant Cell 11:1179–1190

    Article  CAS  PubMed  Google Scholar 

  • Desplats C, Mus F, Cuiné S, Billon E, Cournac L, Peltier G (2009) Characterization of Nda2, a plastoquinone-reducing type II NAD(P)H dehydrogenase in Chlamydomonas chloroplasts. J Biol Chem 284:4148–4157

    Article  CAS  PubMed  Google Scholar 

  • Doebbe A, Rupprecht J, Beckmann J, Mussgnug JH, Hallmann A, Hankamer B, Kruse O (2007) Functional integration of the HUP1 hexose symporter gene into the genome of C. reinhardtii: impacts on biological H2 production. J Biotechnol 131:27–33

    Article  CAS  PubMed  Google Scholar 

  • Edman K, Ericson I, Möller IM (1985) The regulation of exogenous NAD(P)H oxidation in spinach (Spinacia oleracea) leaf mitochondria by pH and cations. Biochem J 232:471–477

    CAS  PubMed  Google Scholar 

  • Erbes DL, King D, Gibbs M (1979) Inactivation of hydrogenase in cell-free extracts and whole cells of Chlamydomonas reinhardi by oxygen. Plant Physiol 63:1138–1142

    Article  CAS  PubMed  Google Scholar 

  • Finazzi G (2002) Redox-coupled proton pumping activity in cytochrome b6f, as evidenced by the pH dependence of electron transfer in whole cells of Chlamydomonas reinhardtii. Biochemistry 41:7475–7482

    Article  CAS  PubMed  Google Scholar 

  • Finazzi G, Rappaport F, Furia A, Fleischmann M, Rochaix J, Zito F, Forti G (2002) Involvement of state transitions in the switch between linear and cyclic electron flow in Chlamydomonas reinhardtii. EMBO Rep 3:280–285

    Article  CAS  PubMed  Google Scholar 

  • Florin L, Tsokoglou A, Happe T (2001) A novel type of iron hydrogenase in the green alga Scenedesmus obliquus is linked to the photosynthetic electron transport chain. J Biol Chem 276:6125–6132

    Article  CAS  PubMed  Google Scholar 

  • Forestier M, King P, Zhang L, Posewitz M, Schwarzer S, Happe T, Ghirardi ML, Seibert M (2003) Expression of two [Fe]-hydrogenases in Chlamydomonas reinhardtii under anaerobic conditions. Eur J Biochem 270:2750–2758

    Article  CAS  PubMed  Google Scholar 

  • Fouchard S, Hemschemeier A, Caruana A, Pruvost J, Legrand J, Happe T, Peltier G, Cournac L (2005) Autotrophic and mixotrophic hydrogen photoproduction in sulfur-deprived Chlamydomonas cells. Appl Environ Microb 71:6199–6205

    Article  CAS  Google Scholar 

  • Frenkel AW (1952) Hydrogen evolution by the flagellate green alga Chlamydomonas moewusii. Arch Biochem 38:219–230

    Article  CAS  PubMed  Google Scholar 

  • Frenkel AW, Rieger C (1951) Photoreduction in algae. Nature 167:1030

    Article  CAS  PubMed  Google Scholar 

  • Gaffron H, Rubin J (1942) Fermentative and photchemical production of hydrogen in algae. J Gen Physiol 26:219–240

    Article  CAS  PubMed  Google Scholar 

  • Gonzalez-Ballester D, Pollock SV, Pootakham W, Grossman AR (2008) The central role of a SNRK2 kinase in sulfur deprivation responses. Plant Physiol 147:216–227

    Article  CAS  PubMed  Google Scholar 

  • Grossman A (2000) Acclimation of Chlamydomonas reinhardtii to its nutrient environment. Protist 151:201–224

    Article  CAS  PubMed  Google Scholar 

  • Guan Y, Zhang W, Deng M, Jin M, Yu X (2004) Significant enhancement of photobiological H2 evolution by carbonylcyanide m-chlorophenylhydrazone in the marine green alga Platymonas subcordiformis. Biotechnol Lett 26:1031–1035

    Article  CAS  PubMed  Google Scholar 

  • Happe T, Kaminski A (2002) Differential regulation of the Fe-hydrogenase during anaerobic adaptation in the green alga Chlamydomonas reinhardtii. Eur J Biochem 269:1022–1032

    Article  CAS  PubMed  Google Scholar 

  • Happe T, Naber JD (1993) Isolation, characterization and N-terminal amino acid sequence of hydrogenase from the green alga Chlamydomonas reinhardtii. Eur J Biochem 214:475–481

    Article  CAS  PubMed  Google Scholar 

  • Happe T, Mosler B, Naber JD (1994) Induction, localization and metal content of hydrogenase in the green alga Chlamydomonas reinhardtii. Eur J Biochem 222:769–774

    Article  CAS  PubMed  Google Scholar 

  • Hemschemeier A, Happe T (2005) The exceptional photofermentative hydrogen metabolism of the green alga Chlamydomonas reinhardtii. Biochem Soc Trans 33:39–41

    Article  CAS  PubMed  Google Scholar 

  • Hemschemeier A, Fouchard S, Cournac L, Peltier G, Happe T (2008) Hydrogen production by Chlamydomonas reinhardtii: an elaborate interplay of electron sources and sinks. Planta 227:397–407

    Article  CAS  PubMed  Google Scholar 

  • Irihimovitch V, Stern DB (2006) The sulfur acclimation SAC3 kinase is required for chloroplast transcriptional repression under sulfur limitation in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 103:7911–7916

    Article  CAS  PubMed  Google Scholar 

  • Irihimovitch V, Yehudai-Resheff S (2008) Phosphate and sulfur limitation responses in the chloroplast of Chlamydomonas reinhardtii. FEMS Microbiol Lett 283:1–8

    Article  CAS  PubMed  Google Scholar 

  • Jans F, Mignolet E, Houyoux P, Cardol P, Ghysels B, Cuiné S, Cournac L, Peltier G, Remacle C, Franck F (2008) A type II NAD(P)H dehydrogenase mediates light-independent plastoquinone reduction in the chloroplast of Chlamydomonas. Proc Natl Acad Sci USA 105:20546–20551

    Article  CAS  PubMed  Google Scholar 

  • Kaltwasser H, Gaffron H (1964) Effects of carbon dioxide and glucose on photohydrogen production in Scenedesmus. Plant Physiol 39(Suppl):xiii

    Google Scholar 

  • Kosourov SN, Seibert M (2009) Hydrogen photoproduction by nutrient-deprived Chlamydomonas reinhardtii cells immobilized within thin alginate films under aerobic and anaerobic conditions. Biotechnol Bioeng 102:50–58

    Article  CAS  PubMed  Google Scholar 

  • Kosourov S, Seibert M, Ghirardi ML (2003) Effects of extracellular pH on the metabolic pathways in sulfur-deprived, H2-producing Chlamydomonas reinhardtii cultures. Plant Cell Physiol 44:146–155

    Article  CAS  PubMed  Google Scholar 

  • Kosourov S, Patrusheva E, Ghirardi ML, Seibert M, Tsygankov A (2007) A comparison of hydrogen photoproduction by sulfur-deprived Chlamydomonas reinhardtii under different growth conditions. J Biotechnol 128:776–787

    Article  CAS  PubMed  Google Scholar 

  • Kruse O (2005) Improved photobiological H2 production in engineered green algal cells. J Biol Chem 280:34170–34177

    Article  CAS  PubMed  Google Scholar 

  • Laurinavichene TV, Kosourov SN, Ghirardi ML, Seibert M, Tsygankov AA (2008) Prolongation of H2 photoproduction by immobilized, sulfur-limited Chlamydomonas reinhardtii cultures. J Biotechnol 134:275–277

    CAS  PubMed  Google Scholar 

  • Lee J, Greenbaum E (2003) A new oxygen sensitivity and its potential application in photosynthetic H2 production. Appl Biochem Biotechnol 106:303–313

    Article  Google Scholar 

  • Melis A (2007) Photosynthetic H2 metabolism in Chlamydomonas reinhardtii (unicellular green algae). Planta 226:1075–1086

    Article  CAS  PubMed  Google Scholar 

  • Melis A, Zhang L, Forestier M, Ghirardi ML, Seibert M (2000) Sustained photobiological hydrogen gas production upon reversible inactivation of oxygen evolution in the green alga Chlamydomonas reinhardtii. Plant Physiol 122:127–136

    Article  CAS  PubMed  Google Scholar 

  • Meuser JE, Ananyev G, Wittig LE, Kosourov S, Ghirardi ML, Seibert M, Dismukes GC, Posewitz MC (2009) Phenotypic diversity of hydrogen production in chlorophycean algae reflects distinct anaerobic metabolisms. J Biotechnol 142:21–30

    Article  CAS  PubMed  Google Scholar 

  • Meyer J (2007) [FeFe] hydrogenases and their evolution: a genomic perspective. Cell Mol Life Sci 64:1063–1084

    Article  CAS  PubMed  Google Scholar 

  • Moreno J, García-Murria MJ, Marín-Navarro J (2008) Redox modulation of Rubisco conformation and activity through its cysteine residues. J Exp Bot 59:1605–1614

    Article  CAS  PubMed  Google Scholar 

  • Ott T, van Dongen JT, Günther C, Krusell L, Desbrosses G, Vigeolas H, Bock V, Czechowski T, Geigenberger P, Udvardi MK (2005) Symbiotic leghemoglobins are crucial for nitrogen fixation in legume root nodules but not for general plant growth and development. Curr Biol 15:531–535

    Article  CAS  PubMed  Google Scholar 

  • Parry MAJ, Keys AJ, Madgwick PJ, Carmo-Silva AE, Andralojc PJ (2007) Rubisco regulation: a role for inhibitors. J Exp Bot 59:1569–1580

    Article  Google Scholar 

  • Posewitz MC, Smolinski SL, Kanakagiri S, Melis A, Seibert M, Ghirardi ML (2004) Hydrogen photoproduction is attenuated by disruption of an isoamylase gene in Chlamydomonas reinhardtii. Plant Cell 16:2151–2163

    Article  CAS  PubMed  Google Scholar 

  • Rühle T, Hemschemeier A, Melis A, Happe T (2008) A novel screening protocol for the isolation of hydrogen producing Chlamydomonas reinhardtii strains. BMC Plant Biol 8:107

    Article  PubMed  Google Scholar 

  • Schönfeld C, Wobbe L, Borgstädt R, Kienast A, Nixon P, Kruse O (2004) The nucleus-encoded protein MOC1 is essential for mitochondrial light acclimation in Chlamydomonas reinhardtii. J Biol Chem 279:50366–50374

    Article  PubMed  Google Scholar 

  • Skjånes K, Knutsen G, Källqvist T, Lindblad P (2008) H2 production from marine and freshwater species of green algae during sulfur deprivation and considerations for bioreactor design. Int J Hydrogen Energy 33:511–521

    Article  Google Scholar 

  • Spruit CJP (1954) Photoproduction of hydrogen and oxygen in Chlorella. In: Proceeding of the First International Photobiology Congress, Amsterdam, pp 323–327

  • Takahashi S, Murata N (2005) Interruption of the Calvin cycle inhibits the repair of Photosystem II from photodamage. Biochim Biophys Acta 1708:352–361

    Article  CAS  PubMed  Google Scholar 

  • Takahashi S, Murata N (2006) Glycerate-3-phosphate, produced by CO2 fixation in the Calvin cycle, is critical for the synthesis of the D1 protein of photosystem II. Biochim Biophys Acta 1757:198–205

    Article  CAS  PubMed  Google Scholar 

  • Timmins M, Thomas-Hall SR, Darling A, Zhang E, Hankamer B, Marx UC, Schenk PM (2009a) Phylogenetic and molecular analysis of hydrogen-producing green algae. J Exp Bot 60:1691–1702

    Article  CAS  PubMed  Google Scholar 

  • Timmins M, Zhou W, Rupprecht J, Lim L, Thomas-Hall SR, Doebbe A, Kruse O, Hankamer B, Marx UC, Smith SM, Schenk PM (2009b) The metabolome of Chlamydomonas reinhardtii following induction of anaerobic H2 production by sulfur depletion. J Biol Chem 284:23415–23425

    Article  Google Scholar 

  • Tolstygina IV, Antal TK, Kosourov SN, Krendeleva TE, Rubin AB, Tsygankov AA (2009) Hydrogen production by photoautotrophic sulfur-deprived Chlamydomonas reinhardtii pre-grown and incubated under high light. Biotechnol Bioeng 102:1055–1061

    Article  CAS  PubMed  Google Scholar 

  • Tsygankov AA, Kosourov SN, Tolstygina IV, Ghirardi ML, Seibert M (2006) Hydrogen production by sulfur-deprived Chlamydomonas reinhardtii under photoautotrophic conditions. Int J Hydrogen Energy 31:1574–1584

    Article  CAS  Google Scholar 

  • Wang ZT, Ullrich N, Joo S, Waffenschmidt S, Goodenough U (2009) Algal lipid bodies: stress induction, purification, and biochemical characterization in wild-type and starchless Chlamydomonas reinhardtii. Eukaryotic Cell 8:1856–1868

    Article  CAS  PubMed  Google Scholar 

  • White AL, Melis A (2006) Biochemistry of hydrogen metabolism in Chlamydomonas reinhardtii wild type and a Rubisco-less mutant. Int J Hydrogen Energy 31:455–464

    Article  CAS  Google Scholar 

  • Winkler M, Heil B, Heil B, Happe T (2002) Isolation and molecular characterization of the [Fe]-hydrogenase from the unicellular green alga Chlorella fusca. Biochim Biophys Acta 1576:330–334

    CAS  PubMed  Google Scholar 

  • Winkler M, Kuhlgert S, Hippler M, Happe T (2009) Characterization of the key steps of light driven hydrogen production in Chlamydomonas reinhardtii. J Biol Chem 284:36620–36627

    Article  CAS  PubMed  Google Scholar 

  • Wollman F-A (2001) State transitions reveal the dynamics and flexibility of the photosynthetic apparatus. EMBO J 20:3623–3630

    Article  CAS  PubMed  Google Scholar 

  • Wu S, Huang R, Xu L, Yan G, Wang Q (2010) Improved hydrogen production with expression of hemH and lba genes in chloroplast of Chlamydomonas reinhardtii. J Biotechnol 146:120–125

    Article  CAS  PubMed  Google Scholar 

  • Wykoff DD, Davies JP, Melis A, Grossman AR (1998) The regulation of photosynthetic electron transport during nutrient deprivation in Chlamydomonas reinhardtii. Plant Physiol 117:129–139

    Article  CAS  PubMed  Google Scholar 

  • Zhang L, Melis A (2002) Probing green algal hydrogen production. Philos Trans R Soc Lond B Biol Sci 357:1499–1511

    Article  CAS  PubMed  Google Scholar 

  • Zhang L, Happe T, Melis A (2002) Biochemical and morphological characterization of sulfur-deprived and H2-producing Chlamydomonas reinhardtii (green alga). Planta 214:552–561

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study has been financed by Action de Recherche Concertée ARC07/12-04. F.F. is a Senior Research Associate of the Fonds de la Recherche Scientifique, F.R.S-FNRS.

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Correspondence to Bart Ghysels.

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Ghysels, B., Franck, F. Hydrogen photo-evolution upon S deprivation stepwise: an illustration of microalgal photosynthetic and metabolic flexibility and a step stone for future biotechnological methods of renewable H2 production. Photosynth Res 106, 145–154 (2010). https://doi.org/10.1007/s11120-010-9582-4

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