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High Photobiological Hydrogen Production Activity of a Nostoc sp. PCC 7422 Uptake Hydrogenase-Deficient Mutant with High Nitrogenase Activity

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Abstract

We describe a strategy to establish cyanobacterial strains with high levels of H2 production that involves the identification of promising wild-type strains followed by optimization of the selected strains using genetic engineering. Nostoc sp. PCC 7422 was chosen from 12 other heterocystous strains, because it has the highest nitrogenase activity. We sequenced the uptake hydrogenase (Hup) gene cluster as well as the bidirectional hydrogenase gene cluster from the strain, and constructed a mutant (ΔhupL) by insertional disruption of the hupL gene. The ΔhupL mutant produced H2 at 100 μmoles mg chlorophyll a -1 h-1, a rate three times that of the wild-type. The ΔhupL cells could accumulate H2 to about 29% (v/v) accompanied by O2 evolution in 6 days, under a starting gas phase of Ar + 5% CO2. The presence of 20% O2 in the initial gas phase inhibited H2 accumulation of the ΔhupL cells by less than 20% until day 7.

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References

  • R Axelsson P Lindblad (2002) ArticleTitleTranscriptional regulation of Nostoc hydrogenases: effect of oxygen, hydrogen and nickel Appl Environ Microbiol 68 444–447 Occurrence Handle10.1128/AEM.68.1.444-447.2002

    Article  Google Scholar 

  • TA Black YP Cai CP Wolk (1993) ArticleTitleSpatial expression and autoregulation of hetR, a gene involved in the control of heterocyst development in Anabaena Mol Microbiol 9 77–84 Occurrence Handle10.1111/j.1365-2958.1993.tb01670.x

    Article  Google Scholar 

  • H Böhme (1998) ArticleTitleRegulation of nitrogen fixation in hetrocyst-forming cyanobacteria Trends Plant Sci 3 346–351 Occurrence Handle10.1016/S1360-1385(98)01290-4

    Article  Google Scholar 

  • G Boison H Bothe O Schmitz (2000) ArticleTitleTranscriptional analysis of hydrogenase genes in the cynobacteria Anacystis nidulans and Anabaena variabilis monitored by RT-PCR Curr Microbiol 40 315–321 Occurrence Handle10.1007/s002849910063

    Article  Google Scholar 

  • YP Cai CP Wolk (1990) ArticleTitleUse of a conditionally lethal gene in Anabaena sp. strain PCC 7120 to select for double recombinants and to entrap insertion sequences J Bacteriol 172 3138–3145

    Google Scholar 

  • CD Carrasco JA Buettner JW Golden (1995) ArticleTitleProgrammed DNA rearrangement of a cyanobacterial hupL gene in heterocysts Proc Natl Acad Sci USA 92 791–795 Occurrence Handle10.1073/pnas.92.3.791

    Article  Google Scholar 

  • Dutta D, De D, Chaudhuri S, Bhattacharya SK (2005) Hydrogen production by cyanobacteria Microbial Cell Factories 4: Art No. 36

    Google Scholar 

  • J Elhai CP Wolk (1988) ArticleTitleConjugal transfer of DNA to cyanobacteria Methods Enzymol 167 747–754 Occurrence Handle10.1016/0076-6879(88)67086-8

    Article  Google Scholar 

  • J Elhai A Vepritskiy AM MuroPastor E Flores CP Wolk (1997) ArticleTitleReduction of conjugal transfer efficiency by three restriction activities of Anabaena sp. strain PCC 7120 J Bacteriol 179 1998–2005

    Google Scholar 

  • A Ernst H Kirschenlohr J Diez P Böger (1984) ArticleTitleGlycogen-content and nitrogenase activity in Anabaena-variabilis Arch Microbiol 140 120–125 Occurrence Handle10.1007/BF00454913

    Article  Google Scholar 

  • DO Hall SA Markov Y Watanabe K Rao (1995) ArticleTitleThe potential applications of cyanobacterial photosynthesis for clean technologies Photosyn Res 46 159–167 Occurrence Handle10.1007/BF00020426

    Article  Google Scholar 

  • A Hansel P Lindblad (1998) ArticleTitleTowards optimization of cyanobacteria as biotechnologically relevant producers of molecular hydrogen, a clean and renewable energy source Appl Microbiol Biotechnol 50 153–160 Occurrence Handle10.1007/s002530051270

    Article  Google Scholar 

  • T Happe K Schutz H Böhme (2000) ArticleTitleTranscriptional and mutational analysis of the uptake hydrogenase of the filamentous cyanobacterium Anabaena variabilis ATCC 29413 J Bacteriol 182 1624–1631 Occurrence Handle10.1128/JB.182.6.1624-1631.2000

    Article  Google Scholar 

  • A Herrero AM Muro-Pastor E Flores (2001) ArticleTitleNitrogen control in cyanobacteria J Bacteriol 183 411–425 Occurrence Handle10.1128/JB.183.2.411-425.2001

    Article  Google Scholar 

  • JC Hwang CH Chen RH Burris (1973) ArticleTitleInhibition of nitrogenase-catalyzed reductions Biochim Biophys Acta 292 256–270 Occurrence Handle10.1016/0005-2728(73)90270-3

    Article  Google Scholar 

  • T Kaneko Y Nakamura CP Wolk T Kuritz S Sasamoto A Watanabe M Iriguchi A Ishikawa K Kawashima T Kimura Y Kishida M Kohara M Matsumoto A Matsuno A Muraki N Nakazaki S Shimpo M Sugimoto M Takazawa M Yamada M Yasuda S Tabata (2001) ArticleTitleComplete genomic sequence of the filamentous nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120 DNA Res 8 227–253 Occurrence Handle10.1093/dnares/8.5.227

    Article  Google Scholar 

  • S Kumazawa H Asakawa (1995) ArticleTitleSimultaneous production of H2 and O2 in closed vessels by marine cyanobacterium Anabaena sp. TU37-1 under high-cell-density conditions Biotechnol Bioengineer 46 396–398 Occurrence Handle10.1002/bit.260460414

    Article  Google Scholar 

  • S Kumazawa A Mitsui (1994) ArticleTitleEfficient hydrogen photoproduction by synchronously grown cells of a marine cyanobacterium, Synechococcus sp Miami BG 043511, under high cell density conditions Biotechnol Bioengineer 44 854–858 Occurrence Handle10.1002/bit.260440711

    Article  Google Scholar 

  • P Lindberg A Hansel P Lindblad (2000) ArticleTitle hupS and hupL constitute a transcription unit in the cyanobacterium Nostoc sp. PCC 73102 Arch Microbiol 174 129–133 Occurrence Handle10.1007/s002030000186

    Article  Google Scholar 

  • P Lindberg K Schütz T Happe P Lindblad (2002) ArticleTitleA hydrogen-producing, hydrogenase-free mutant strain of Nostoc punctiforme ATCC 29133 Intern J Hydrogen Energy 27 1291–1296 Occurrence Handle10.1016/S0360-3199(02)00121-0

    Article  Google Scholar 

  • P Lindberg F Lindblad L Cournac (2004) ArticleTitleGas exchange in the filamentous cyanobacterium Nostoc punctiforme strain ATCC 29133 and its hydrogenase-deficient mutant strain NHM5 Appl Environ Microbiol 70 2137–2145 Occurrence Handle10.1128/AEM.70.4.2137-2145.2004

    Article  Google Scholar 

  • H Masukawa K Nakamura M Mochimaru H Sakurai (2001) “Photobiological hydrogen production and nitrogenase activity in some heterocystous cyanobacteria” J Miyake T Matsunaga A San Pietro (Eds) Biohydrogen II Elsevier Amsterdam 63–66

    Google Scholar 

  • H Masukawa M Mochimaru H Sakurai (2002a) ArticleTitleDisruption of the uptake hydrogenase gene, but not of the bidirectional hydrogenase gene, leads to enhanced photobiological hydrogen production by nitrogen-fixing cyanobacterium Anabaena sp. PCC 7120 Appl Microbiol Biotechnol 58 618–624 Occurrence Handle10.1007/s00253-002-0934-7

    Article  Google Scholar 

  • H Masukawa M Mochimaru H Sakurai (2002b) ArticleTitleHydrogenases and photobiological hydrogen production utilizing nitrogenase system in cyanobacteria Int J Hydrogen Energy 27 1471–1474 Occurrence Handle10.1016/S0360-3199(02)00125-8

    Article  Google Scholar 

  • LE Mikheeva O Schmitz SV Shestakov H Bothe (1995) ArticleTitleMutants of the cyanobacterium Anabaena variabilis altered in hydrogenase activities Z Naturforsch 50c 505–510

    Google Scholar 

  • FAL Pinto O Troshina P Lindblad (2002) ArticleTitleA brief look at three decades of research on cyanobacterial hydrogen evolution Intern J Hydrogen Energy 27 1209–1215 Occurrence Handle10.1016/S0360-3199(02)00089-7

    Article  Google Scholar 

  • RC Prince HS Kheshgi (2005) ArticleTitleThe photobiological production of hydrogen: potential efficiency and effectiveness as a renewable fuel Critical Rev Microbiol 31 19–31 Occurrence Handle10.1080/10408410590912961

    Article  Google Scholar 

  • J Reaston GC Duyvesteyn A Waard Particlede (1982) ArticleTitle Nostoc PCC7524, a cyanobacteria which contains five sequence-specific deoxyribonucleases Gene 20 103–110 Occurrence Handle10.1016/0378-1119(82)90091-9

    Article  Google Scholar 

  • R Rippka M Herdman (1992) Pasteur Culture Collection of Cyanobacterial Strains in Axenic Culture, Vol. I: Catalogue of Strains Institut Pasteur Paris

    Google Scholar 

  • H Sakurai H Masukawa S Dawar F Yoshino (2004) “Photobiological hydrogen production by cyanobacteria utilizing nitrogenase systems–Present status and future development” J Miyake Y Igarashi M Rögner (Eds) Biohydrogen III Elsevier Amsterdam 84–93

    Google Scholar 

  • J Sambrook EF Fritsch T Maniatis (1989) Molecular Cloning: A Laboratory Manual EditionNumber2nd ed Cold Spring Harbor Laboratory Press Cold Harbor, NY

    Google Scholar 

  • O Schmitz G Boison H Salzmann H Bothe K Schutz S Wang T Happe (2002) ArticleTitleHoxE — a subunit specific for the pentameric bidirectional hydrogenase complex (HoxEFUYH) of cyanobacteria Biochim Biophys Acta 1554 66–74 Occurrence Handle10.1016/S0005-2728(02)00214-1

    Article  Google Scholar 

  • K Schütz T Happe O Troshina P Lindblad E Leitao P Oliveira P Tamagnini (2004) ArticleTitleCyanobacterial H2 production–a comparative analysis Planta 218 350–359 Occurrence Handle10.1007/s00425-003-1113-5

    Article  Google Scholar 

  • L Serebriakova NA Zorin P Lindblad (1994) ArticleTitleReversible hydrogenase in Anabaena variabilis ATCC 29413–presence and localization in non-N2-fixing cells Arch Microbiol 161 140–144

    Google Scholar 

  • S Spiro (1994) ArticleTitleThe FNR family of transcriptional regulators Antonie Van Leeuwenhoek. Int J Gen Mol Microbiol 66 23–36 Occurrence Handle10.1007/BF00871630

    Article  Google Scholar 

  • WDP Stewart HW Pearson (1970) ArticleTitleEffects of aerobic and anaerobic conditions on growth and metabolism of blue-green algae Proc Roy Soc 175B 293–311

    Google Scholar 

  • P Tamagnini R Axelsson P Lindberg F Oxelfelt R Wünschiers P Lindblad (2002) ArticleTitleHydrogenases and hydrogen metabolism of cyanobacteria Microbiol Mol Biol Rev 66 1–20 Occurrence Handle10.1128/MMBR.66.1.1-20.2002

    Article  Google Scholar 

  • CM Thomas CA Smith (1987) ArticleTitleIncompatibility group P plasmids: genetics, evolution, and use in genetic manipulation Annu Rev Microbiol 41 77–101 Occurrence Handle10.1146/annurev.mi.41.100187.000453

    Article  Google Scholar 

  • PM Vignais A Colbeau (2004) ArticleTitleMolecular biology of microbial hydrogenases Curr Issues Mol Biol 6 159–188

    Google Scholar 

  • PM Vignais B Billoud J Meyer (2001) ArticleTitleClassification and phylogeny of hydrogenases FEMS Microbiol Rev 25 455–501

    Google Scholar 

  • R Wagner (2000) Transcription Regulation in Prokaryotes Oxford University Press Oxford

    Google Scholar 

  • Z-C Wang GD Watt (1984) ArticleTitleH2-uptake activity of the MoFe protein component of Azotobacter vinelandii nitrogenase Proc Natl Acad Sci USA 81 376–379 Occurrence Handle10.1073/pnas.81.2.376

    Article  Google Scholar 

Download references

Acknowledgments

We thank C. P. Wolk for the pRL plasmids. This work was supported in part by the MEXT grant Venture (2001–2005), the MEXT grant Initiatives for Attractive Education for Graduate Schools (No b043), Grants for Special Research Projects, Waseda University (2004A-085, 2005B-083) to HS and Grants-in-Aid for Scientific Research from JSPS (16.9494) to HM.

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Correspondence to Hidehiro Sakurai.

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Yoshino, F., Ikeda, H., Masukawa, H. et al. High Photobiological Hydrogen Production Activity of a Nostoc sp. PCC 7422 Uptake Hydrogenase-Deficient Mutant with High Nitrogenase Activity. Mar Biotechnol 9, 101–112 (2007). https://doi.org/10.1007/s10126-006-6035-3

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