Photosynthesis Research

, 80:401 | Cite as

Trails of Green Alga Hydrogen Research – from Hans Gaffron to New Frontiers

  • Anastasios Melis
  • Thomas Happe


This paper summarizes aspects of the history of photosynthetic hydrogen research, from the pioneering discovery of Hans Gaffron over 60 years ago to the potential exploitation of green algae in commercial H2-production. The trail started as a mere scientific curiosity, but promises to be a most important discovery, one that leads photosynthesis research to important commercial applications. Progress achieved in the field of photosynthetic hydrogen production by green algae includes elucidation of the mechanism, the ability to modify photosynthesis by physiological means and to produce bulk amounts of H2 gas, and cloning of the [Fe]-hydrogenase genes in several green algal species.

Hans Gaffron green algae hydrogen photosynthesis 


  1. Adams MWW (1990) The structure and mechanism of ironhydrogenases. Biochim Biophys Acta 1020: 115–145PubMedCrossRefGoogle Scholar
  2. Adams MWW and Stiefel EI (2000) Organometallic iron: the key to biological hydrogen metabolism. Curr Opin Chem Biol 4: 214–220PubMedCrossRefGoogle Scholar
  3. Arnon DI, Mitsui A and Paneque A (1961) Photoproduction of hydrogen gas coupled with photosynthetic phosphorylation. Science 134: 1425–1425Google Scholar
  4. Bamberger ES, King D, Erbes DL and Gibbs M (1982) H2 and CO2evolution by anaerobically adapted Chlamydomonas reinhardtii F60. Plant Physiol 69: 1268–1273PubMedGoogle Scholar
  5. Ben-Amotz A and Avron M(1990) The biotechnology of cultivating the halotolerant alga Dunaliella. Trends Biotechnol 8: 121–128CrossRefGoogle Scholar
  6. Ben-Amotz A, Erbes DL, Riederer-Henderson MA, Peavey DG and Gibbs M (1975) H2 metabolism in photosythetic organism. I. Dark H2 evolution and uptake by algae and mosses. Plant Physiol 56: 72–77PubMedGoogle Scholar
  7. Benemann JR (1996) Hydrogen biotechnology: progress and prospects. Nat Biotechnol 14: 1101–1103PubMedCrossRefGoogle Scholar
  8. Bennoun P (2001) Chlororespiration and the process of carotenoid biosynthesis. Biochim Biophys Acta 1506: 133–142PubMedCrossRefGoogle Scholar
  9. Bennoun P (2002) The present model for chlororespiration. Photosynth Res 73: 273–277PubMedCrossRefGoogle Scholar
  10. Bishop NI (1966) Partial reactions of photosynthesis and photoreduction. Ann Rev Plant Physiol 17: 185–208CrossRefGoogle Scholar
  11. Bishop NI and Gaffron H (1963) On the interrelation of the mechanisms for oxygen and hydrogen evolution in adapted algae. In: Kok B and Jagendorf AT (eds) Photosynthetic Mechanisms in Green Plants, Publ 1145, pp 441–451. Natl Acad Sci Natl Res Council, Washington, DCGoogle Scholar
  12. Bishop NI, Frick M and Jones LW (1977) Photohydrogen production in green algae: water serves as the primary substrate for hydrogen and oxygen production. In: Mitsui A, Miyachi S, San Pietro A and Tamura S (eds) Biological Solar Energy Conversion, pp 3–22. Academic Press, New YorkGoogle Scholar
  13. Cinco RM, Macinnis JM and Greenbaum E (1993) The role of carbon dioxide in light-activated hydrogen production by Chlamydomonas reinhardtii. Photosynth Res 38: 27–33CrossRefGoogle Scholar
  14. Erbes DL, King D and Gibbs M (1979) Inactivation of hydrogenase in cell-free extracts and whole cells of Chlamydomonas reinhardtii by oxygen. Plant Physiol 63: 1138–1142PubMedGoogle Scholar
  15. Feild TS, Nedbal L and Ort DR (1998) Nonphotochemical reduction of the plastoquinone pool in sunflower leaves originates from chlororespiration. Plant Physiol 116: 1209–1218PubMedCrossRefGoogle Scholar
  16. Florin L, Tsokoglou A and Happe T (2001) A novel type of [Fe]-hydrogenase in the green alga Scenedesmus obliquus is linked to the photosynthetical electron transport chain. J Biol Chem 276: 6125–6132PubMedCrossRefGoogle Scholar
  17. Francis K and Senger H (1985) Correlation betweeen respiration and hydrogenase adaptation in Scenedesmus obliquus. Physiol Plant 65: 167–170CrossRefGoogle Scholar
  18. Frenkel AW (1952) Hydrogen evolution of the flagellate green alga Chlamydomonas moewusi. Arch Biochem Biophys 38: 219–230PubMedCrossRefGoogle Scholar
  19. Frenkel AW and Lewin RA (1954) Photoreduction by Chlamydomonas. Am J Bot 41: 586–589CrossRefGoogle Scholar
  20. Gaffron H (1939) Reduction of CO2 with H2 in green plants. Nature 143: 204–205Google Scholar
  21. Gaffron H (1940) Carbon dioxide reduction with molecular hydrogen in green algae. Am J Bot 27: 273–283CrossRefGoogle Scholar
  22. Gaffron H (1942) Reduction of carbon dioxide coupled with the oxyhydrogen reaction in algae. J Gen Physiol 26: 241–267CrossRefPubMedGoogle Scholar
  23. Gaffron H (1944) Photosynthesis, photoreduction and dark reduction of carbon dioxide in certain algae. Biol Rev Cambridge Phil Soc 19: 1–20CrossRefGoogle Scholar
  24. Gaffron H and Rubin J (1942) Fermentative and photochemical production of hydrogen in algae. J Gen Physiol 26: 219–240CrossRefPubMedGoogle Scholar
  25. Gfeller RP and Gibbs M (1984) Fermentative metabolism of Chlamydomonas reinhardtii. I. Analysis of fermentative products from starch in dark-light. Plant Physiol 75: 212–218PubMedGoogle Scholar
  26. Gfeller RP and Gibbs M (1985) Fermentative metabolism of Chlamydomonas reinhardtii. II. Role of plastoquinone. Plant Physiol 77: 509–511PubMedGoogle Scholar
  27. Ghirardi ML, Togasaki RK and Seibert M(1997) Oxygen sensitivity of algal H2-production. Appl Biochem Biotech 63: 141–151Google Scholar
  28. Ghirardi ML, Zhang L, Lee JW, Flynn T, Seibert M, Greenbaum E and Melis A (2000) Microalgae: a green source of renewable H2.Trends Biotechnol 18: 506–511PubMedCrossRefGoogle Scholar
  29. Gibbs M, Gfeller RP and Chen C (1986) Fermentative metabolism of Chlamydomonas reinhardtii. III. Photo-assimilation of acetate. Plant Physiol 82: 160–166PubMedGoogle Scholar
  30. Godde D and Trebst A (1980) NADH as electron donor for the photosynthetic membrane of Chlamydomonas reinhardtii. Arch Microbiol 127: 245–252CrossRefGoogle Scholar
  31. Greenbaum E (1982) Photosynthetic hydrogen and oxygen production: kinetic studies. Science 196: 879–880Google Scholar
  32. Greenbaum E (1988) Energetic efficiency of H2 photoevolution by algal water-splitting. Biophys J 54: 365–368Google Scholar
  33. Greenbaum E, Guillard RRL and Sunda WG (1983) Hydrogen and oxygen photoproduction by marine algae. Photochem Photobiol 37: 649–655Google Scholar
  34. Happe T and Kaminski A (2002) Differential regulation of the [Fe]-hydrogenase during anaerobic adaptation in the green alga Chlamydomonas reinhardtii. Eur J Biochem 269: 1022–1032PubMedCrossRefGoogle Scholar
  35. Happe T and Naber JD (1993) Isolation, characterization and Nterminal amino acid sequence of hydrogenase from the green alga Chlamydomonas reinhardtii. Eur J Biochem 214: 475–481PubMedCrossRefGoogle Scholar
  36. Happe T, Mosler B and Naber JD (1994) Induction, localization and metal content of hydrogenase in the green alga Chlamydomonas reinhardtii. Eur J Biochem 222: 769–774PubMedCrossRefGoogle Scholar
  37. Happe T, Hemschemeier A, Winkler M and Kaminski A (2002) Hydrogenases in green algae: do they safe the algae's life and solve our energy problems? Trends Plant Sci 7: 246–250PubMedCrossRefGoogle Scholar
  38. Healey FP (1970) The mechanism of hydrogen evolution by Chlamydomonas moewusii. Plant Physiol 45: 153–159PubMedGoogle Scholar
  39. Homann PH (2003) Hydrogen metabolism of green algae. Discovery and early research — a tribute to Hans Gaffron and his coworkers. Photosynth Res 76: 93–103PubMedCrossRefGoogle Scholar
  40. Horner DS, Heil B, Happe T and Embley TM (2002) Iron hydrogenases, ancient enzymes in modern eukaryotes. Trends Biochem Sci 27: 148–153PubMedCrossRefGoogle Scholar
  41. Kaltwasser H, Stuart TS and Gaffron H (1969) Light-dependent hydrogen evolution by Scenedesmus. Planta 89: 309–322CrossRefGoogle Scholar
  42. Kessler E (1966) The effect of glucose on hydrogenase activity in Chlorella. Biochim Biophys Acta 112: 173–175PubMedGoogle Scholar
  43. Kessler E (1973) Effect of anaerobiosis on photosynthetic reactions and nitrogen metabolism of algae with and without hydrogenase. Arch Microbiol 93: 91–100Google Scholar
  44. Kessler E (1974) Hydrogenase, photoreduction and anaerobic growth of algae. In: Stewart WDP (ed) Algal Physiology and Biochemistry, pp 454–473. Blackwell, OxfordGoogle Scholar
  45. Klein U and Betz A (1978) Fermentative metabolism of hydrogenevolving Chlamydomonas moewusii. Plant Physiol 61: 953–956PubMedGoogle Scholar
  46. Lien S and San Pietro S (1981) Effect of uncouplers on anaerobic adaptation of hydrogenase activity in C. reinhardtii. Biochem Biophys Res Comm 103: 139–147PubMedCrossRefGoogle Scholar
  47. Maione TE and Gibbs M (1986a) Association of the chloroplastic respiratory and photosynthetic electron transport chains of C. reinhardii with photoreduction and the oxyhydrogen reaction. Plant Physiol 80: 364–368PubMedGoogle Scholar
  48. Maione TE and Gibbs M (1986b) Hydrogenase-mediated activities in isolated chloroplasts of Chlamydomonas reinhardii. Plant Physiol 80: 360–363PubMedGoogle Scholar
  49. McBride AC, Lien S, Togasaki RK and San Pietro A (1977) Mutational analysis of Chlamydomonas reinhardi: application to biological solar energy conversion. In: Mitsui A, Miyachi S, San Pietro A and Tamura S (eds) Biological Solar Energy Conversion, pp 77–86. Academic Press, New YorkGoogle Scholar
  50. Melis A and Happe T (2001) Hydrogen production: green algae as a source of energy. Plant Physiol 127: 740–748PubMedCrossRefGoogle Scholar
  51. Melis A, Zhang L, Forestier M, Ghirardi ML and Seibert M (2000) Sustained photobiological hydrogen gas production upon reversible inactivation of oxygen evolution in the green alga Chlamydomonas reinhardtii. Plant Physiol 122: 127–136PubMedCrossRefGoogle Scholar
  52. Miura Y (1995) Hydrogen production by biophotolysis based on microalgal photosynthesis. Process Biochem 30: 1–7CrossRefGoogle Scholar
  53. Miura Y, Ohta S, Mano M and Miyamoto K (1986) Isolation and characterization of a unicellular green alga exhibiting high activity in dark hydrogen production. Agric Biol Chem 50: 2837–2844Google Scholar
  54. Miura Y, Saitoh C, Matsuoka S and Miyamoto K (1992) Stably sustained hydrogen production with high molar yield through a combination of a marine green alga and a photosynthetic bacterium. Biosci Biotechnol Biochem 56: 751–754CrossRefGoogle Scholar
  55. Miyaki J, Matsunaga T and San Pietro A (2001) BioHydorgen II. An Approach to Environmentally Acceptable Technology, pp 1–273. Pergamon Press, New YorkGoogle Scholar
  56. Miyamoto K, Nawa Y, Matsuoka S, Ohta S and Miura Y (1990) Mechanism of adaptation and hydrogen photoproduction in a marine green alga Chlamydomonas sp. MGA 161. J Ferment Bioeng 70: 66–69CrossRefGoogle Scholar
  57. Nicolet Y, Lemon BJ, Fontecilla-Camps JC and Peters JW(2000) A novel FeS cluster in Fe-only hydrogenases. Trends Biochem Sci 25: 138–142PubMedCrossRefGoogle Scholar
  58. Nicolet Y, de Lacey AL, Vernede X, Fernandez VM, Hatchikian EC and Fontecilla-Camps JC (2001) Crystallographic and FTIR spectroscopic evidence of changes in Fe coordination upon reduction of the active site of the Fe-only hydrogenase from Desulfovibrio desulfuricans. J Am Chem Soc 123: 1596–1601PubMedCrossRefGoogle Scholar
  59. Ohta S, Miyamoto K and Miura Y (1987) Hydrogen evolution as a consumption mode of reducing equivalents in green alga fermentation. Plant Physiol 83: 1022–1026PubMedCrossRefGoogle Scholar
  60. Peters JW (1999) Structure and mechanism of iron-only hydrogenases. Curr Opin Struct Biol 9: 670–676PubMedCrossRefGoogle Scholar
  61. Peters JW, Lanzilotta WN, Lemon BJ and Seefeldt LC (1998) Xray crystal structure of the [Fe]-only hydrogenase (CpI) from Clostridium pasteurianum to 1.8 Angstrom resolution. Science 282: 1853–1858PubMedCrossRefGoogle Scholar
  62. Randt C and Senger H (1985) Participation of the two photosystems in light dependent hydrogen evolution in Scenedesmus obliquus. Photochem Photobiol 42: 553–557Google Scholar
  63. Roessler PG and Lien S (1984a) Activation and de novo synthesis of hydrogenase in Chlamydomonas. Plant Physiol 76: 1086–1089PubMedGoogle Scholar
  64. Roessler PG and Lien S (1984b) Purification of hydrogenase from Chlamydomonas reinhardtii. Plant Physiol 75: 705–709PubMedGoogle Scholar
  65. Schulz R (1996) Hydrogenases and hydrogen production in eukaryotic organisms and cyanobacteria. J Mar Biotechnol 4: 16–22Google Scholar
  66. Smith BM, Morrissey PJ, Guenther JE, Nemson JA, Harrison MA, Allen JF and Melis A (1990) Response of the photosynthetic apparatus in Dunaliella salina (green algae) to irradiance stress. Plant Physiol 93: 1433–1440PubMedGoogle Scholar
  67. Spruit CP (1958) Simultaneous photoproduction of hydrogen and oxygen by Chlorella. Meded Landbouwhogesch Wageningen 58: 1–17Google Scholar
  68. Stuart TS and Gaffron H (1971) The kinetics of hydrogen photoproduction by adapted Scenedesmus. Planta (Berlin) 100: 228–243Google Scholar
  69. Stuart TS and Gaffron H (1972a) The mechanism of hydrogen photoproduction by several algae. I. The effect of inhibitors of photophosphorylation. Planta (Berlin) 106: 91–100Google Scholar
  70. Stuart TS and Gaffron H (1972b) The mechanism of hydrogen photoproduction by several algae. II. The contribution of Photosystem II. Planta (Berlin) 106: 101–112Google Scholar
  71. Van Neil EWJ, Janssen M, Lindblad P, Barten H, Reith JH and Wijffels RH (2002) BioHydrogen 2002 (Special Issue). Int J Hydrogen Energy 27: 1123–1505CrossRefGoogle Scholar
  72. Vignais PN, Billoud B and Meyer J (2001) Classification and phylogeny of hydrogenases. FEMS Microbiol Rev 25: 455–501PubMedGoogle Scholar
  73. Winkler M, Heil B and Happe T (2002) Isolation and molecular characterization of the [Fe]-hydrogenase from the unicellular green alga Chlorella fusca. Biochim Biophys Acta 1576: 330–334PubMedGoogle Scholar
  74. Wunschiers R, Stangier K, Senger H and Schulz R (2001) Molecular evidence for a [Fe]-hydrogenase in the green alga Scenedesmus obliquus. Curr Microbiol 42: 353–360PubMedCrossRefGoogle Scholar
  75. Wykoff DD, Davies JP, Melis A and Grossman AR (1998) The regulation of photosynthetic electron-transport during nutrient deprivation in Chlamydomonas reinhardtii. Plant Physiol 117: 129–139PubMedCrossRefGoogle Scholar
  76. Zaborski OR (1998) Biohydrogen, pp 1–552. Plenum Press, New YorkGoogle Scholar
  77. Zhang L, Happe T and Melis A (2002) Biochemical and morphological characterization of sulfur-deprived and H2-producing Chlamydomonas reinhardtii (green alga). Planta 214: 552–561PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Anastasios Melis
    • 1
  • Thomas Happe
    • 2
  1. 1.Department of Plant and Microbial BiologyUniversity of CaliforniaBerkeleyUSA
  2. 2.Biochemie der Pflanzen, AG PhotobiotechnologieRuhr-Universität-BochumBochumGermany

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