Journal of Applied Phycology

, Volume 25, Issue 2, pp 575–585 | Cite as

Factors affecting biohydrogen production by unicellular halotolerant cyanobacterium Aphanothece halophytica

Article

Abstract

The effects of several physiological parameters on H2 production rate in the unicellular halotolerant cyanobacterium Aphanothece halophytica were investigated. Under nitrogen deprivation, the growth of cells was inhibited, but H2 production rate was enhanced approximately fourfold. Interestingly, cells grown under sulfur deprivation exhibited a decrease in cell growth, H2 production rate, and bidirectional hydrogenase activity. Glucose was the preferred sugar source for H2 production by A. halophytica, but H2 production decreased at high glucose concentrations. H2 production rate was optimum when cells were grown in the presence of 0.75 M NaCl, or 0.4 μM Fe3+, or 1 μM Ni2+. The optimum light intensity and temperature for H2 production were 30 μmol photons m−2 s−1 and 35 °C, respectively. A two-stage culture of A. halophytica was performed in order to overcome the reduction of cell growth in N-free medium. In the first stage, cells were grown in normal medium to accumulate biomass, and in the second stage, H2 production by the obtained biomass was induced by growing cells in N-free medium supplemented with various chemicals for 24 h. A. halophytica grown in N-free medium containing various MgSO4 concentrations had a high H2 production rate between 11.432 and 12.767 μmol H2 mg chlorophyll a (chl a)−1 h−1, a 30-fold increase compared to cells grown in normal medium. The highest rate of 13.804 μmol H2 mg chl a−1 h−1 was obtained when the N-free growth medium contained 0.4 μM Fe3+. These results suggested the possibility of using A. halophytica and some other halotolerant cyanobacteria thriving under extreme environmental conditions in the sea as potential sources for H2 production in the future.

Keywords

Hydrogen production Cyanobacteria Aphanothece halophytica Two-stage culture 

References

  1. Ananyev G, Carrieri D, Dismukes GC (2008) Optimization of metabolic capacity and flux through environmental cues to maximize hydrogen production by the cyanobacterium “Arthrospira maxima”. Appl Environ Microbiol 74:6102–6113PubMedCrossRefGoogle Scholar
  2. Antal TK, Lindblad P (2005) Production of H2 by sulphur-deprived cells of the unicellular cyanobacteria Gloeocapsa alpicola and Synechocystis sp. PCC 6803 during dark incubation with methane or at various extracellular pH. J Appl Microbiol 98:114–120PubMedCrossRefGoogle Scholar
  3. Axelsson R, Lindblad P (2002) Transcriptional regulation of Nostoc hydrogenases: effects of oxygen, hydrogen, and nickel. Appl Environ Microbiol 68:444–447PubMedCrossRefGoogle Scholar
  4. Baebprasert W, Lindblad P, Incharoensakdi A (2010) Response of H2 production and Hox-hydrogenase activity to external factors in the unicellular cyanobacterium Synechocystis sp. strain PCC 6803. Int J Hydrog Energy 35:6611–6616CrossRefGoogle Scholar
  5. Baebprasert W, Jantaro S, Khetkorn W, Lindblad P, Incharoensakdi A (2011) Increased H2 production in the cyanobacterium Synechocystis sp. strain PCC 6803 by redirecting the electron supply via genetic engineering of the nitrate assimilation pathway. Metab Eng 13:610–616PubMedCrossRefGoogle Scholar
  6. Belkin S, Padan E (1978) Hydrogen metabolism in the facultative anoxygenic cyanobacteria (Blue-green algae) Oscillatoria limnetica and Aphanothece halophytica. Arch Microbiol 116:109–111PubMedCrossRefGoogle Scholar
  7. Carrieri D, Ananyev G, Garcia Costas AM, Bryant DA, Dismukes GC (2008) Renewable hydrogen production by cyanobacteria: nickel requirements for optimal hydrogenase activity. Int J Hydrog Energy 33:2014–2022CrossRefGoogle Scholar
  8. Chen PC, Fan SH, Chiang CL, Lee CM (2008) Effect of growth conditions on the hydrogen production with cyanobacterium Anabaena sp. strain CH3. Int J Hydrog Energy 33:1460–1464CrossRefGoogle Scholar
  9. Cournac L, Guedeney G, Peltier G, Vignais PS (2004) Sustained photoevolution of molecular hydrogen in a mutant of Synechocystis sp. strain PCC 6803 deficient in the type I NADPH-dehydrogenase complex. J Bacteriol 186:1737–1746PubMedCrossRefGoogle Scholar
  10. Daday A, Mackerras AH, Smith GD (1985) The effect of nickel on hydrogen metabolism and nitrogen fixation in the cyanobacterium Anabaena cylindrica. J Gen Microbiol 131:231–238Google Scholar
  11. Dawar S, Mohanty P, Behera BK (1999) Sustainable hydrogen production in the cyanobacterium Nostoc sp. ARM 411 grown in fructose- and magnesium sulphate-enriched culture. World J Microbiol Biotechnol 15:329–332CrossRefGoogle Scholar
  12. Dean DR, Bolin JT, Zheng L (1993) Nitrogen metalloclusters: structure, organization, and synthesis. J Bacteriol 175:6737–6744PubMedGoogle Scholar
  13. Dutta D, De D, Chaudhuri S, Bhattacharya SK (2005) Hydrogen production by cyanobacteria. Microb Cell Fact 4:36–46PubMedCrossRefGoogle Scholar
  14. Ferreira D, Stal LJ, Moradas-Ferreira P, Mendes MV, Tamagnini P (2009) The relation between N2 fixation and H2 metabolism in the marine filamentous nonheterocystous cyanobacterium Lyngbya aestuarii CYY 9616. J Phycol 45:898–905CrossRefGoogle Scholar
  15. Garlick S, Oren A, Padan E (1977) Occurrence of facultative anoxygenic photosynthesis among filamentous and unicellular cyanobacteria. J Bacteriol 29:623–629Google Scholar
  16. Geider RJ, Roche J (1994) The role of iron in phytophankton photosynthesis, and the potential for iron limitation of primary productivity in the sea. Photosynth Res 399:275–301CrossRefGoogle Scholar
  17. Gutekunst K, Hoffmann D, Lommer M, Egert M, Suzuki I, Schulz-Friedrich R, Appel J (2006) Metal dependence and intracellular regulation of the bidirectional NiFe-hydrogenase in Synechocystis sp. PCC 6803. Int J Hydrog Energy 31:1452–1459CrossRefGoogle Scholar
  18. Hibino T, Kaku N, Yoshikawa H, Takabe T, Takabe T (1999) Molecular characterization of DnaK from the halotolerant cyanobacterium Aphanothece halophytica for ATPase, protein folding, and copper binding under various salinity conditions. Plant Mol Biol 40:409–418PubMedCrossRefGoogle Scholar
  19. Incharoensakdi A, Waditee-Sirisattha R (2013) Regulatory mechanisms of cyanobacteria in response to osmotic stress. In: Srivastava et al. (eds) Stress biology of cyanobacteria: molecular mechanisms to cellular responses. Taylor & Francis/CRC Press, Boca Raton, Florida (in press)Google Scholar
  20. Ishitani M, Takabe T, Kojima K, Takabe T (1993) Regulation of glycinebetaine accumulation in the halotolerant cyanobacterium Aphanothece halophytica. Aust J Plant Physiol 20:693–703CrossRefGoogle Scholar
  21. Jeffries TW, Timourian H, Ward RL (1978) Hydrogen production by Anabaena cylindrica: effect of varying ammonium and ferric ions, pH and light. Appl Environ Microbiol 35:704–710PubMedGoogle Scholar
  22. Khetkorn W, Lindblad P, Incharoensakdi A (2010) Enhanced biohydrogen production by the N2-fixing cyanobacterium Anabaena siamensis strain TISTR 8012. Int J Hydrog Energy 35:12767–12776CrossRefGoogle Scholar
  23. Khetkorn W, Baebprasert W, Lindblad P, Incharoensakdi A (2012) Redirecting the electron flow towards the nitrogenase and bidirectional Hox-hydrogenase by using specific inhibitors results in enhanced H2 production in the cyanobacterium Anabaena siamensis TISTR 8012. Bioresour Technol 118:265–271PubMedCrossRefGoogle Scholar
  24. Kumar S, Polasa H (1991) Influence of nickel and copper on photobiological hydrogen production and uptake in Oscillatoria subbrevis strain111. Proc Indian Natl Sci Acad B57:281–285Google Scholar
  25. Kumazawa S (2003) Photoproduction of hydrogen by the marine heterocystous cyanobacterium Anabaena species TU37-1 under a nitrogen atmosphere. Mar Biotechnol 5:222–226PubMedCrossRefGoogle Scholar
  26. Kumazawa S, Asakawa H (1995) Simultaneous production of H2 and O2 in closed vessels by marine cyanobacterium Anabaena sp. TU37-1 under high-cell-density conditions. Biotechnol Bioeng 46:396–398PubMedCrossRefGoogle Scholar
  27. Kumazawa S, Mitsui A (1981) Characterization and optimization of hydrogen photoproduction by saltwater blue-green algae, Oscillatoria sp. Miami BG7. I. Enhancement through limiting the supply of nitrogen nutrients. Int J Hydrog Energy 6:339–348CrossRefGoogle Scholar
  28. Kumazawa S, Mitsui A (1994) Efficient hydrogen photoproduction by synchronous grown cell of a marine cyanobacterium, Synechococcus sp. Miami BG 043511, under high cell density conditions. Biotechnol Bioeng 44:854–858PubMedCrossRefGoogle Scholar
  29. Kumazawa S, Shimamura K (1993) Photosynthesis-dependent production of H2 by a marine cyanobacterium, Anabaena sp. TU37-1. J Mar Biotechnol 1:159–162Google Scholar
  30. Kuwada Y, Ohta Y (1989) Hydrogen production and carbohydrate consumption by Lyngbya sp. (No. 108). Agric Biol Chem 53:2847–2851CrossRefGoogle Scholar
  31. Lambert GR, Smith GD (1977) Hydrogen formation by marine blue-green algae. FEBS Lett 83:159–162PubMedCrossRefGoogle Scholar
  32. Lin JT, Stewart V (1997) Nitrate assimilation by bacteria. Adv Microb Physiol 39:1–30CrossRefGoogle Scholar
  33. Ludwig M, Schulz-Friedrich R, Appel J (2006) Occurrence of hydrogenases in cyanobacteria and anoxygenic photosynthetic bacteria: implications for the phylogenetic origin of cyanobacterial and algal hydrogenases. J Mol Evol 63:758–768PubMedCrossRefGoogle Scholar
  34. Luo YH, Mitsui A (1994) Hydrogen production from organic substrates in an aerobic nitrogen-fixing marine unicellular cyanobacterium Synechococcus sp. strain Miami BG 043511. J Biotechnol Bioeng 44:1255–1260CrossRefGoogle Scholar
  35. MacKinney G (1941) Absorption of light by chlorophyll solutions. J Biol Chem 140:315–322Google Scholar
  36. Melis A, Zhang LP, 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 122:127–135PubMedCrossRefGoogle Scholar
  37. Min H, Sherman LA (2010) Hydrogen production by the unicellular, diazotroph cyanobacterium Cyanothece sp. strain ATCC 51142 under conditions of continuous light. Appl Environ Microbiol 76:4293–4301PubMedCrossRefGoogle Scholar
  38. Oxelfelt F, Tamagnini P, Salema R, Lindblad P (1995) Hydrogen uptake in Nostoc strain PCC73102: effects of nickel, hydrogen, carbon and nitrogen. Plant Physiol Biochem 33:617–623Google Scholar
  39. Perry JH (1963) Chemical engineers’ handbook. McGraw-Hill, New YorkGoogle Scholar
  40. Phlips EJ, Mitsui A (1983) Role of light intensity and temperature in the regulation of hydrogen photoproduction by the marine cyanobacterium Oscillatoria sp. strain Miami BG7. Appl Environ Microbiol 45:1212–1220PubMedGoogle Scholar
  41. Pinto FL, Troshina O, Lindblad P (2002) A brief look at three decades of research on cyanobacterial hydrogen evolution. Int J Hydrog Energy 27:1209–1215CrossRefGoogle Scholar
  42. Prabaharan D, Subramanian G (1996) Oxygen-free hydrogen production by the marine cyanobacterium Phormidium valderianum BDU 20041. Bioresour Technol 57:111–116CrossRefGoogle Scholar
  43. Prabaharan D, Arun-Kumar D, Uma L, Subramanian G (2010) Dark hydrogen production in nitrogen atmosphere—an approach for sustainability by marine cyanobacterium Leptolyngbya valderiana BDU 20041. Int J Hydrog Energy 35:10725–10730CrossRefGoogle Scholar
  44. Prince RC, Kheshgi HS (2005) The photobiological production of hydrogen: potential efficiency and effectiveness as a renewable fuel. Crit Rev Microbiol 31:19–31PubMedCrossRefGoogle Scholar
  45. Rai LC, Raizada M (1986) Nickel induced stimulation of growth, heterocyst differentiation, 14CO2 uptake and nitrogenase activity in Nostoc muscorum. New Phytol 104:111–114CrossRefGoogle Scholar
  46. Rashid N, Song W, Park J, Jin H-F, Lee K (2009) Characteristics of hydrogen production by immobilized cyanobactrium Microcystis aeruginosa through cycles of photosynthesis and anaerobic incubation. J Ind Eng Chem 15:498–503CrossRefGoogle Scholar
  47. Raven JA, Evans MCW, Korbs RE (1999) The role of trace metals in photosynthetic electron transport in O2-evolving organisms. Photosynth Res 60:111–149CrossRefGoogle Scholar
  48. Reddy PM, Spiller H, Albrecht SL, Shanmugam KT (1996) Photodissimilation of fructose to H2 and CO2 by a dinitrogen-fixing cyanobacterium, Anabaena variabilis. Appl Environ Microbiol 62:1220–1226PubMedGoogle Scholar
  49. Serebryakova LT, Sheremetieva M, Tsygankov AA (1998) Reversible hydrogenase activity of Gloeocapsa alpicola in continuous culture. FEMS Microbiol Lett 166:89–94CrossRefGoogle Scholar
  50. Serebryakova LT, Sheremetieva M, Lindblad P (1999) Hydrogenase activity of the unicellular cyanobacterium Gloeocapsa alpicola CALU743 under conditions of nitrogen limitation. Microbiology 68:249–253Google Scholar
  51. Shah V, Garg N, Madamwar D (2001) Ultrastructure of the fresh water cyanobacterium Anabaena variabilis SPU 003 and its application for oxygen-free hydrogen production. FEMS Microbiol Lett 194:71–75PubMedCrossRefGoogle Scholar
  52. Shah V, Garg N, Madamwar D (2003) Ultrastructure of the cyanobacterium Nostoc muscorum and exploitation of the culture for hydrogen production. Folia Microbiol 48:65–70CrossRefGoogle Scholar
  53. Suda S, Kumazawa S, Mitsui A (1992) Change in the H2 photoproduction capability in a synchronously grown aerobic nitrogen-fixing cyanobacterium, Synechococcus sp. Miami BG 043511. Arch Microbiol 158:1–4CrossRefGoogle Scholar
  54. Takabe T, Incharoensakdi A, Arakawa K, Yokota S (1988) CO2 fixation rate and RuBisCO content increase in the halotolerant cyanobacterium, Aphanothece halophytica, grown in high salinities. Plant Physiol 88:1120–1124PubMedCrossRefGoogle Scholar
  55. Tel-Or E, Melhamed-Harel H (1981) Adaptation to salt of photosynthetic apparatus in cyanobacteria. In: Akoyunoglou G (ed) Photosynthesis. Bablan International Science Services, Philadelphia, pp 455–462Google Scholar
  56. Tindall DR, Yopp JH, Miller DM, Schmid WE (1978) Physico-chemical parameters governing the growth of Aphanothece halophytica (Chroococcales) in hypersaline media. Phycologia 17:179–185CrossRefGoogle Scholar
  57. Troshina O, Serebryakova LT, Sheremetieva M, Lindblad P (2002) Production of H2 by the unicellular cyanobacterium Gloeocapsa alpicola CALU743 during fermentation. Int J Hydrog Energy 27:1283–1289CrossRefGoogle Scholar
  58. Vignais PM, Billoud B, Meyer J (2001) Classification and phylogeny of hydrogenases. FEMS Microbiol Rev 25:455–501PubMedGoogle Scholar
  59. Weissman JC, Benemann JR (1977) Hydrogen production by nitrogen-starved cultures of Anabaena cylindrica. Appl Env Microbiol 33:123–131Google Scholar
  60. Wilson ST, Tozzi S, Foster RA et al (2010) Hydrogen cycling by the unicellular marine diazotroph Crocosphaera watsonii strain WH8501. Appl Environ Microbiol 76:6797–6803PubMedCrossRefGoogle Scholar
  61. Wutipraditkul N, Waditee R, Incharoensakdi A et al (2005) Halotolerant cyanobacterium Aphanothece halophytica contains NapA-type Na+/H+ antiporters with novel ion specificity that are involved in salt tolerance at alkaline pH. Appl Environ Microbiol 71:4176–4184PubMedCrossRefGoogle Scholar
  62. Xiankong Z, Haskell JB, Tabita FR, Van Baalen C (1983) Aerobic hydrogen production by the heterocystous cyanobacteria Anabaena spp. strains CA and 1F. J Bacteriol 156:1118–1122Google Scholar
  63. Xiankong Z, Tabita FR, Van Baalen C (1984) Nickel control of hydrogen production and uptake in Anabaena spp. strains CA and 1F. J Gen Microbiol 130:1815–1818Google Scholar
  64. Zhang L, Happe T, Melis A (2002) Biochemical and morphological characterization of sulphur-deprived and H2-producing Chlamydomonas reinhardtii (green alga). Planta 214:552–561PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  1. 1.Department of Biology, Faculty of ScienceKing Mongkut’s Institute of Technology LadkrabangBangkokThailand
  2. 2.Department of Chemistry, Faculty of ScienceKing Mongkut’s Institute of Technology LadkrabangBangkokThailand
  3. 3.Laboratory of Cyanobacterial Biotechnology, Department of Biochemistry, Faculty of ScienceChulalongkorn UniversityBangkokThailand

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