Applied Microbiology and Biotechnology

, Volume 92, Issue 6, pp 1161–1169 | Cite as

Cell growth and P(3HB) accumulation from CO2 of a carbon monoxide-tolerant hydrogen-oxidizing bacterium, Ideonella sp. O-1

  • Kenji TanakaEmail author
  • Kenta Miyawaki
  • Akane Yamaguchi
  • Kianoush Khosravi-Darani
  • Hiromi Matsusaki
Biotechnological Products and Process Engineering


Cell growth and accumulation of polyhydroxybutyric acid, P(3HB), from CO2 in autotrophic condition of a newly isolated hydrogen-oxidizing bacterium, the strain O-1, was investigated. The bacterium, which was deposited in the Japan Collection of Microorganisms as JCM17105, autotrophically grows by assimilating H2, O2, and CO2 as substrate. 16S rRNA gene sequence of the bacterium was the closest to Ideonella dechloratans (99%). Specific growth rate of the strain O-1 was faster than a hydrogen-oxidizing bacterium, Ralstonia eutropha, which is well-known P(3HB)-producing microorganism. The strain O-1 is tolerant to high O2 concentration and it can grow above 30% (v/v) O2, while the growth of R. eutropha and Alcaligenes latus was seriously inhibited. In culture medium containing 1 g/L (NH4)2SO4, cell concentration of the strain O-1 and P(3HB) increased to 6.75 and 5.26 g/L, respectively. The content of P(3HB) in the cells was 77.9% (w/w). The strain O-1 was very tolerant to carbon monoxide (CO) and it grew even at 70% (v/v) CO, while the growth of R. eutropha and A. latus were seriously inhibited at 5% (v/v) CO. From these results, it is expected that the strain O-1 will be useful in the manufacture of P(3HB) because the industrial exhaust gas containing CO2, H2, and CO can be directly used as the substrate in the fermentation process.


Hydrogen-oxidizing bacteria Ideonella PHB CO2 CO 



A part of this study was supported by a scientific grant from JFE Steel Corporation, Japan.


  1. Ahn WS, Park SJ, Lee SY (2000) Production of poly(3-hydroxybutyrate) by fed-batch culture of recombinant Escherichia coli with a highly concentrated whey solution. Appl Environ Microbiol 66:3624–3627CrossRefGoogle Scholar
  2. Ammann ECB, Reed LL, Durichek JE Jr (1968) Gas consumption and growth rate of Hydrogenomonas eutropha in continuous culture. Appl Microbiol 16:822–826Google Scholar
  3. Anderson A, Dawes EA (1990) Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev 54:450–472Google Scholar
  4. Bongers L (1970) Energy generation and utilization in hydrogen bacteria. J Bacteriol 104:145–151Google Scholar
  5. Byrom D (1994) Polyhydroxyalkanoates. In: Mobley DP (ed) Plastics from microbes: microbial synthesis of polymers and polymer precursors. Hanser, Munich, pp 5–33Google Scholar
  6. Choi J, Lee SY (1999a) Factors affecting the economics of polyhydroxyalkanoate production by bacterial fermentation. Appl Microbiol Biotechnol 51:13–21CrossRefGoogle Scholar
  7. Choi J, Lee SY (1999b) High-level production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by fed-batch culture of recombinant Escherichia coli. Appl Environ Microbiol 65:4346–4348Google Scholar
  8. Elbeltagy A, Nishioka K, Sato T, Suzuki H, Ye B, Hamada T, Isawa T, Mitsui H, Minamisawa K (2001) Endophytic colonization and in planta nitrogen fixation by a Herbaspirillum sp. isolated from wild rice species. Appl Environ Microbiol 67:5285–5293CrossRefGoogle Scholar
  9. Goto E, Suzuki K, Kodama T (1977) Improvement of initial and exponential growth of hydrogen bacteria, strain 9-5. Agr Biol Chem 41:521–525CrossRefGoogle Scholar
  10. Ishizaki A, Tanaka K (1990) Batch culture of Alcaligenes eutrophus ATCC 17697T using recycled gas closed circuit culture system. J Ferment Bioeng 69:170–174CrossRefGoogle Scholar
  11. Ishizaki A, Tanaka K (1991) Production of poly-β-hydroxybutyric acid from carbon dioxide by Alcaligenes eutrophus ATCC 17697T. J Ferment Bioeng 70:254–257CrossRefGoogle Scholar
  12. Kim BS, Lee SC, Lee SY, Chang HN, Chang YK, Woo SI (1994) Production of poly(3-hydroxybutyric-co-3-hydroxyvaleric acid) by fed-batch culture of Alcaligenes eutrophus with substrate control using on-line glucose analyzer. Enzyme Microbiol Technol 16:556–561CrossRefGoogle Scholar
  13. Kodama T, Igarashi Y, Minoda Y (1975) Isolation and culture conditions of a bacterium grown on hydrogen and carbon dioxide. Agr Biol Chem 39:77–82CrossRefGoogle Scholar
  14. Lee IY, Kim MK, Kim GK, Chang HN, Park YH (1995) Production of poly(β-hydroxybutyrate-co-β-hydroxyvalerate) from glucose and valerate in Alcaligenes eutrophus. Biotechnol Lett 17:571–574CrossRefGoogle Scholar
  15. Lee SY, Choi J, Wong HH (1999) Recent advances in polyhydroxyalkanoate production by bacterial fermentation. Int J Biol Macromol 25:31–36CrossRefGoogle Scholar
  16. Malmqvist A, Welander T, Moore E, Ternström A, Molin G, Stenström IM (1994) Ideonella dechloratans gen. nov., sp. nov., a new bacterium capable of growing anaerobically with chlorate as an electron acceptor. Syst Appl Microbiol 17:58–64Google Scholar
  17. Miura Y, Okazaki M, Ohi K, Nishimura T, Komemushi S (1982) Optimization of biomass productivity and substrate utility of a hydrogen bacterium, Alcaligenes hydrogenophilus. Biotechnol Bioeng 24:1173–1182CrossRefGoogle Scholar
  18. Morinaga Y, Yamanaka S, Ishizaki A, Hirose Y (1978) Growth characteristics and cell composition of Alcaligenes eutrophus in chemostat culture. Agr Biol Chem 42:439–444CrossRefGoogle Scholar
  19. Noar JD, Buckley DH (2009) Ideonella azoifingens sp. nov., an aerobic diazotroph of the Betaproteobacteria isolated from grass rhizosphere soil, and emended description of the genus Ideonella. Int J Syst Evol Microbiol 59:1941–1946CrossRefGoogle Scholar
  20. Ohi K, Matsumoto H, Yamada K (1979) Cultivation and characterization of a hydrogen bacterium, Alcaligenes hydrogenophilus sp. nov. J Ferment Technol 57:195–202Google Scholar
  21. Repaske R (1961) Nutritional requirements for Hydrogenomonas eutropha. J Bacteriol 83:418–422Google Scholar
  22. Repaske R (1966) Characteristics of hydrogen bacteria. Biotechnol Bioeng 8:217–235CrossRefGoogle Scholar
  23. Repaske R, Mayer R (1976) Dense autotrophic cultures of Alcaligenes eutrophus. Appl Environ Microbiol 32:592–597Google Scholar
  24. Repaske R, Repaske AC (1976) Quantitative requirement for exponential growth of Alcaligenes eutrophus. Appl Environ Microbiol 32:585–591Google Scholar
  25. Ryu HW, Hann SK, Chang YK, Chang HN (1997) Production of poly(3-hydroxyvutyrate) by high cell density fed-batch culture of Alcaligenes eutrophus with phosphate limitation. Biotechnol Bioeng 55:28–32CrossRefGoogle Scholar
  26. Savelieva ND (1979) Microbiology. On response of hydrogen bacteria to carbon monoxide. Mikrobiologiya 48:360–362 (in Russian)Google Scholar
  27. Savelieva ND, Zhilina TN (1968) On systematics of hydrogen bacteria. Mikrobiologiya 37:84–91 (in Russian)Google Scholar
  28. Schlegel HG (1989) Aerobic hydrogen-oxidizing (Knallgas) bacteria. In: Schlegel HG, Bowien B (eds) Autotrophic bacteria. Science Tech, Madison, pp 305–329Google Scholar
  29. Schlegel HG, Lafferty RM (1971) Novel energy and carbon sources. The production of biomass from hydrogen and carbon dioxide. Adv Biochem Eng 143:143–168CrossRefGoogle Scholar
  30. Sonnleitner B, Heinzle E, Braunegg G, Lafferty RM (1979) Formal kinetics of poly-β-hydroxybutyric acid (PHB) production in Alcaligenes eutrophus H16 and Mycoplana rubra R14 with respect to the dissolved oxygen tension in ammonium-limited batch cultures. Eur J Appl Microbiol Biotechnol 7:1–10CrossRefGoogle Scholar
  31. Stasishina GN, Volova TG (1996) The strain of bacteria Alcaligenes eutrophus. RF Patent No. 2053292. Bull. No. 3Google Scholar
  32. Steinbüchel A, Hein S (2001) Biochemical and molecular basis of microbial synthesis of polyhydroxyalkanoates in microorganisms. Adv Biochem Eng Biotechnol 71:81–123Google Scholar
  33. Sugimoto T, Tsuge T, Tanaka K, Ishizaki A (1999) Control of acetic acid concentration by pH-stat continuous substrate feeding in heterotrophic culture phase of two-stage cultivation of Alcaligenes eutrophus for production of P(3HB) from CO2, H2 and O2 under non-explosive condition. Biotechnol Bioeng 62:625–631CrossRefGoogle Scholar
  34. Suzuki T, Yamane T, Shimizu S (1986) Mass production of poly-β-hydroxybutyric acid by fed-batch culture with controlled carbon/nitrogen feeding. Appl Microbiol Biotechnol 24:370–374CrossRefGoogle Scholar
  35. Taga N, Tanaka K, Ishizaki A (1997) Effects of rheological change by addition of carboxymethylcellulose in culture media of an air-lift fermentor on poly-d-3-hydroxybutyric acid productivity in autotrophic culture of hydrogen-oxidizing bacterium, Alcaligenes eutrophus. Biotechnol Bioeng 53:529–533CrossRefGoogle Scholar
  36. Takeshita T, Tanaka K, Ishizaki A, Stanbury PF (1993) Development of a dissolved hydrogen sensor and its application to evaluation of hydrogen mass transfer. J Ferment Bioeng 76:148–150CrossRefGoogle Scholar
  37. Tanaka K, Ishizaki A (1994) Production of poly-D-3-hydroxybutyric acid from carbon dioxide by a two-stage culture method employing Alcaligenes eutrophus ATCC 17697T. J Ferment Bioeng 77:425–427CrossRefGoogle Scholar
  38. Tanaka K, Ishizaki A, Kanamaru T, Kawano T (1995) Production of poly(D-3-hydroxybutyrate) from CO2, H2, and CO2 by high cell density autotrophic cultivation of Alcaligenes eutrophus. Biotechnol Bioeng 45:268–275CrossRefGoogle Scholar
  39. Volova TG, Kalacheva GS, Stasishina GN, Kasaeva GE (1980) Investigation of the growth of hydrogen bacteria under inhibition by carbon monoxide. Mikrobiologiya 49:465–471 (in Russian)Google Scholar
  40. Volova TG, Guseinov OA, Kalacheva GM, Medvedeva SE, Puzyr AAP (1993) Effect of carbon monoxide on metabolism and ultrastructure of carboxydobacteria. World J Microbiol Biotechnol 9:160–163CrossRefGoogle Scholar
  41. Volova TG, Kalacheva GS, Altukhova OV (2002) Autotrophic synthesis of polyhydroxyalkanoates by the bacteria Ralstonia eutropha in the presence of carbon monoxide. Appl Microbiol Biotechnol 58:675–678CrossRefGoogle Scholar
  42. Wang F, Lee SY (1997a) Production of poly(3-hydroxybutyrate) by fed-batch culture of filamentation-suppressed recombinant Escherichia coli. Appl Environ Microbiol 63:4765–4769Google Scholar
  43. Wang F, Lee SY (1997b) Poly(3-hydroxybutyrate) production of with high productivity and high polymer content by a fed-batch culture of Alcaligenes latus under nitrogen limitation. Appl Environ Microbiol 63:3703–3706Google Scholar
  44. Yamane T, Fukunaga M, Lee YW (1996) Increase PHB production by high-cell-density fed-batch culture of Alcaligenes latus, a growth-associated PHB producer. Biotechnol Bioeng 50:197–202CrossRefGoogle Scholar
  45. Zinn M, Witholt B, Egli T (2001) Occurrence, synthesis and medical application of bacterial polyhydroxyalkanoate. Adv Drug Deliv Rev 53(1):5–21CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Kenji Tanaka
    • 1
    Email author
  • Kenta Miyawaki
    • 1
  • Akane Yamaguchi
    • 1
  • Kianoush Khosravi-Darani
    • 2
  • Hiromi Matsusaki
    • 3
  1. 1.Department of Biological and Environmental Chemistry, Faculty of Humanity-Oriented Science and EngineeringKinki UniversityFukuokaJapan
  2. 2.National Nutrition and Food Technology Research InstituteShahid Beheshti Medical UniversityTehranIran
  3. 3.Department of Food and Health Sciences, Faculty of Environmental and Symbiotic SciencesPrefectural University of KumamotoKumamotoJapan

Personalised recommendations