Skip to main content
Log in

Distribution and Selection of Poly-3-Hydroxybutyrate Production Capacity in Methanotrophic Proteobacteria

  • Environmental Microbiology
  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

Methanotrophs are known to produce poly-3-hydroxybutyrate (PHB), but there is conflicting evidence in the literature as to which genera produce the polymer. We screened type I and II proteobacterial methanotrophs that use the ribulose monophosphate and serine pathways for carbon assimilation, respectively, for both phaC, which encodes for PHB synthase, and the ability to produce PHB under nitrogen-limited conditions. Twelve strains from six different genera were evaluated. All type I strains tested negative for phaC and PHB production; all Type II strains tested positive for phaC and PHB production. In order to identify conditions that favor PHB production, we also evaluated a range of selection conditions using a diverse activated sludge inoculum. Use of medium typically recommended for methanotroph enrichment led to enrichments dominated by type I methanotrophs. Conditions that were selected for enrichments dominated by PHB-producing Type II methanotrophs were: (1) use of nitrogen gas as the sole nitrogen source in the absence of copper, (2) use of a dilute mineral salts media in the absence of copper, and (3) use of media prepared at pH values of 4–5.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Amaral JA, Archambault C, Richards SR, Knowles R (1995) Denitrificationassociated with groups I and II methanotrophs in a gradient enrichment system. FEMS Microbiol Ecol 18:289–298

    Article  CAS  Google Scholar 

  2. Anderson AJ, Dawes EA (1990) Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev 54:450–472

    PubMed  CAS  Google Scholar 

  3. Asenjo JA, Suk J (1986) Microbial conversion of methaneintopoly-beta-hydroxybutrate (PHB)-growth and intracellular product accumulation in a type-II methanotroph. J Ferment Technol 64:271–278

    Article  CAS  Google Scholar 

  4. Ashby RD, Solaiman DKY, Foglia TA (2004) Bacterialpoly (hydroxyalkanoate) polymer production from the bio diesel co-product stream. J Polym Environ 12:105–112

    Article  CAS  Google Scholar 

  5. Auman AJ, Speake CC, Lidstrom ME (2001) nifH sequences and nitrogen fixation in type I and type II methanotrophs. Appl Environ Microbiol 67:4009–4016

    Article  PubMed  CAS  Google Scholar 

  6. Babel W (1992) Pecularities of methylotrophs concerning over flow metabolism, especially the synthesis of polyhydroxyalkanoates. FEMS Microbiol Rev 103:141–148

    Article  CAS  Google Scholar 

  7. Bowman JP (2001) Family I. Methylococcaceae and Family V. Methylocystaceae. In: Bergey’s manual of systematic bacteriology. Williams & Wilkins, Baltimore, pp 256–270, pp. 411–420

    Google Scholar 

  8. Bowman JP (2006) The methanotrophs-the families Methylococcacceae and Methylocystaceae. In: The prokaryotes a: handbook on the biology of bacteria. Springer, NewYork, pp 266–289

    Google Scholar 

  9. Bowman JP, Sly LI, Nichols PD, Hayward A (1993) Revised Taxonomy of the Methanotrophs: Description of Methylobacter gen.nov., emendation of Methylococcus, validation of Methylosinus and Methylocystis species, and a proposal that the family Methylococcaceae includes only the group I methanotrophs. Int J Syst Bacteriol 43:735–753

    Article  Google Scholar 

  10. Braunegg G, Lefebvre G, Genser KF (1999) Polyhydroxyalkanoates, biopolyesters from renewable resources: physiological and engineering aspects. J Biotechnol 65:127–161

    Article  Google Scholar 

  11. Braunegg G, Sonnleitner B, Lafferty R (1978) Arapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass. Eur J Appl Microbiol Biotechnol 6:29–37

    Article  CAS  Google Scholar 

  12. Budwill K, Fedorak PM, Page WJ (1992) Methanogenic degradation of poly (3- hydroxyalkanoates). Appl Environ Microbiol 58:1398–1401

    PubMed  CAS  Google Scholar 

  13. Bussman I, Pester M, Brune A, Schink B (2004) Preferential cultivation of type II methanotrophic bacteria from littoral sediments (Lake Constance). FEMS Microbiol Ecol 47:179–189

    Article  Google Scholar 

  14. Cebron A, Bodrossy L, Stralis-Pavese N, Singer AC, Thompson IP, Prosser JI, Murrell JC (2007) Nutrient amendments in soil DNA stable isotope probing experiments reduce the observed methanotroph diversity. Appl Environ Microbiol 73:798–807

    Article  PubMed  CAS  Google Scholar 

  15. Choi J, Lee SY (1997) Process analysis and economic evaluation for Poly (3- hydroxybutyrate) production by fermentation. Bioprocess Eng 17:335–342

    Article  CAS  Google Scholar 

  16. Costello AM, Lidstrom ME (1999) Molecular characterization of functional and phylogenetic genes from natural populations of methanotrophs in lake sediments. Appl Environ Microbiol 65:5066–5074

    PubMed  CAS  Google Scholar 

  17. Dedysh SN (2002) Methanotrophic bacteria of acids phagnum bogs. Mikrobiologiia 71:741–754

    PubMed  CAS  Google Scholar 

  18. Dedysh SN, Khmelenina VN, Suzina NE, Trotsenko YA, Semrau JD, Liesack W, Tiedje JM (2002) Methylocapsa acidiphila gen. nov., sp. nov., a novel methane-oxidizing and dinitrogen-fixing acidophilic bacterium from sphagnum bog. Int J Syst Evol Microbiol 52:251–261

    PubMed  CAS  Google Scholar 

  19. Dedysh SN, Liesack W, Khmelenina VN, Suzina NE, Trotsenko YA, Semrau JD, Bares AM, Panikov NS, Tiedje JM (2000) Methylocellapalustrisgen.nov., sp.nov., a new methane-oxidizing acid ophilic bacterium from peat bogs, representing a novel subtype of serine-pathway methanotrophs. Int J Syst Evol Microbiol 50:955–969

    Article  PubMed  CAS  Google Scholar 

  20. Du G, Chen LXL, Yu J (2004) High-efficiency production of bioplastics from biodegradable organic solids. J Polym Environ 12:89–94

    Article  CAS  Google Scholar 

  21. Dunfield PF, Khmelenina VN, Suzina NE, Trotsenko YA, Dedysh SN (2003) Methylocella silvestrissp.nov.,a novel methanotroph isolated from an acidic forest cambisol. Int J Syst Evol Microbiol 53:1231–1239

    Article  PubMed  CAS  Google Scholar 

  22. Dunfield PF, Yuryev A, Senin P, Smirnova AV, Stott MB, Hou S, Ly B, Saw JH, Zhou Z, Ren Y, Wang J, Mountain BW, Crowe MA, Weatherby TM, Bodelier PL, Liesack W, Feng L, Wang L, Alam M (2007) Methane oxidation by an extremely acid ophilic bacterium of the phylum Verrucomicrobia. Nature 450:879–882

    Article  PubMed  CAS  Google Scholar 

  23. Follner CG, Babel W, Valentin HE, Steinbuchel A (1993) Expression of polyhydroxy alkanoic-acid-biosynthesis genes in methylotrophic bacteria relying on the ribulose monophosphate pathway. Appl Microbiol Biotechnol 40:284–291

    Article  Google Scholar 

  24. Graham DW, Chaudhary JA, Hanson RS, Arnold RG (1993) Factors affecting competition between type I and type II methanotrophs in two-organism, continuous-flowreactors. Microb Ecol 25:1–17

    Article  CAS  Google Scholar 

  25. Hanson RS (1998) Ecology of methylotrophic bacteria. In: Techniques in microbial ecology. Oxford University Press, London, pp 137–162

    Google Scholar 

  26. Hanson RS, Hanson TE (1996) Methanotrophic bacteria. Microbiol Rev 60:439–471

    PubMed  CAS  Google Scholar 

  27. Harrington AA, Kallio RE (1960) Oxidation of methanol and formal dehyde by Pseudomonas methanica. Can J Microbiol 6:1–7

    Article  PubMed  CAS  Google Scholar 

  28. Helm J, Wendlandt KD, Jechorek M, Stottmeister U (2008) Potassium deficiency results in accumulation of ultra-high molecular weight poly-beta-hydroxybutyrate in a methane-utilizing mixed culture. J Appl Microbiol 105:1054–1061

    Article  PubMed  CAS  Google Scholar 

  29. Helm J, Wendlandt KD, Rogge G, Kappelmeyer U (2006) Characterizing a stable methane-utilizing mixed culture used in the synthesis of a high-quality biopolymer in an open system. J Appl Microbiol 101:387–395

    Article  PubMed  CAS  Google Scholar 

  30. Heyer J, Berger U, Hardt M, Dunfield PF (2005) Methylohalobiuscrimeensisgen.nov., sp.nov., a moderately halophilic, methanotrophic bacterium isolated from hyper-saline lakes of Crimea. Int J Syst Evol Microbiol 55:1817–1826

    Article  PubMed  CAS  Google Scholar 

  31. Higgins IJ, Best DJ, Hammond RC, Scott D (1981) Methane-oxidizing microorganisms. Microbiol Rev 45:556–590

    PubMed  CAS  Google Scholar 

  32. Islam T, Jensen S, Reigstad LJ, Larsen O, Birkeland NK (2008) Methane oxidation at 55 degrees C and pH2 by a thermo acid ophilic bacterium belonging to the Verrucomicrobia phylum. Proc Natl Acad Sci USA 105:300–304

    Article  PubMed  CAS  Google Scholar 

  33. Kallio RE, Harrington AA (1960) Sudanophilic granules and lipid of Pseudomonas methanica. J Bacteriol 80:321–324

    PubMed  CAS  Google Scholar 

  34. Koller M, Bona R, Braunegg G, Hermann C, Horvat P, Kroutil M, Martinz J, Neto J, Pereira L, Varila P (2005) Production of polyhydroxyalkanoates from agricultural waste and surplus materials. Biomacro Molecules 6:561–565

    Article  CAS  Google Scholar 

  35. Korotkova N, Lidstrom ME (2001) Connection between poly-beta-hydroxybutyrate biosynthesis and growth on C (1) and C (2) compounds in the methylotroph Methylobacterium extorquens AM1. J Bacteriol 183:1038–1046

    Article  PubMed  CAS  Google Scholar 

  36. Lee SY, Park SJ, Park JP, Lee Y, Lee SH (2005) Economic aspects of biopolymer production, vol 2. WILEY-VCH, Weinheim

    Google Scholar 

  37. Lidstrom ME (2006) Aerobic methylotrophic prokaryotes. In: Prokaryotes. pp. 618–634

  38. Madison LL, Huisman GW (1999) Metabolic engineering of poly (3-hydroxyalkanoates): from DNA to plastic. Microbiol Mol Biol Rev 63:21–53

    PubMed  CAS  Google Scholar 

  39. Morse M, Liao Q, Criddle CS, Frank CW (2011) An aerobic biodegradation of the microbial copolymer poly (3-hydroxybutyrate-co-3-hydroxyhexanoate): Effects of comonomer content, processing history, and semi-crystalline morphology. Polymer 52:547–555

    Article  CAS  Google Scholar 

  40. Murrell J, Dalton H (1983) Nitrogen-fixation in obligate methanotrophs. J Gen Microbiol 129:3481–3486

    CAS  Google Scholar 

  41. Nguyen HH, Elliott SJ, Yip JH, Chan SI (1998) The particulate methane monooxygenase from M. capsulatus (Bath) is a novel copper-containing three-subunit enzyme. J Biol Chem 273:7957–7966

    Article  PubMed  CAS  Google Scholar 

  42. Oakley C, Murrell J (1988) Nifh genes in the obligate methane oxidizing bacteria. FEMS Microbiol Lett 49:53–57

    Article  Google Scholar 

  43. HJ Op denCamp, Islam T, Stott MB, Harhangi HR, Hynes A, Schouten S, Jetten MS, Birkeland NK, Pol A, Dunfield PF (2009) Environmental, genomic and taxonomic perspectives on methanotrophic Verrucomicrobia. Environ Microbiol Rep 1:293–306

    Article  Google Scholar 

  44. Pol A, Heijmans K, Harhangi HR, Tedesco D, Jetten MS, OpdenCamp HJ (2007) Methanotrophy below pH1 by a new Verrucomicrobia species. Nature 450:874–878

    Article  PubMed  CAS  Google Scholar 

  45. Povolo S, Casella S (2003) Bacterial production of PHA from lactose and cheese whey permeate. Macromol Symp 197:1–9

    Article  CAS  Google Scholar 

  46. Shah NN, Hanna ML, Jackson KJ, Taylor RT (1996) Batch cultivation of Methylosinus trichosporium OB3B: IV production of hydrogen-driven soluble or particulate methane monooxygenase activity. Biotechnol Bioeng 45:229–238

    Article  Google Scholar 

  47. Sheu DS, Wang YT, Lee CY (2000) Rapid detection of polyhydroxyalkanoate-accumulating bacteria isolated from the environment by colony PCR. Microbiology 146:2019–2025

    PubMed  CAS  Google Scholar 

  48. Stocks PK, Mc Cleskey CS (1964) Identity of the pink-pigmented methanol- oxidizing bacteria as Vibrio extorquens. J Bacteriol 88:1065–1070

    PubMed  CAS  Google Scholar 

  49. U.S.EnvironmentalProtectionAgency. Methane: sources and emissions. http://www.epa.gov/outreach/sources.html. Accessed April21, 2011

  50. Vecherskaya M, Dijkema C, Stams AJ (2001) Intracellular PHB conversion in a type II methanotroph studied by13CNMR. J Ind Microbiol Biotechnol 26:15–21

    Article  CAS  Google Scholar 

  51. Vorob’ev AV, Dedysh SN (2008) Use of enrichment cultures for assessing the structure of methanotrophic communities in peat soil: the problem of representativity of the results. Mikrobiologiia 77:566–569

    PubMed  Google Scholar 

  52. Ward N, Larsen O, Sakwa J, Bruseth L, Khouri H, Durkin AS, Dimitrov G, Jiang L, Scanlan D, Kang KH, Lewis M, Nelson KE, Methe B, Wu M, Heidelberg JF, Paulsen IT, Fouts D, Ravel J, Tettelin H, Ren Q, Read T, DeBoy RT, Seshadri R, Salzberg SL, Jensen HB, Birkeland NK, Nelson WC, Dodson RJ, Grindhaug SH, Holt I, Eidhammer I, Jonasen I, Vanaken S, Utterback T, Feldblyum TV, Fraser CM, Lillehaug JR, Eisen JA (2004) Genomic insights into methanotrophy: the complete genome sequence of M. capsulatus (Bath). PLoS Biol 2:e303

    Article  PubMed  Google Scholar 

  53. Wendlandt KD, Geyer W, Mirschel G, Al-HajHemidi F (2005) Possibilities for controlling a PHB accumulation process using various analytical methods. J Biotechnol 117:119–129

    Article  PubMed  CAS  Google Scholar 

  54. Wendlandt KD, Jechorek M, Helm J, Stottmeister U (2001) Producing poly-3-hydroxybutyrate with a high molecular mass from methane. J Biotechnol 86:127–133

    Article  PubMed  CAS  Google Scholar 

  55. Wise MG, McArthur JV, Shimkets LJ (1999) Methanotroph diversity in land fill soil: isolation of novel type I and type II methanotrophs whose presence was suggested by culture-independent 16S ribosomal DNA analysis. Appl Environ Microbiol 65:4887–4897

    PubMed  CAS  Google Scholar 

  56. Xin JY, Zhang YX, Zhang S, Xia CG, Li SB (2007) Methanol production from CO2by resting cells of the methanotrophic bacterium Methylosinus trichosporium IMV3011. J Basic Microbiol 47:426–435

    Article  PubMed  CAS  Google Scholar 

  57. Zahn JA, DiSpirito AA (1996) Membrane-associated methane monooxygenase from M. capsulatus (Bath). J Bacteriol 178:1018–1029

    PubMed  CAS  Google Scholar 

  58. Zhang Y, Xin J, Chen L, Song H, Xia C (2008) Biosynthesis of poly-3-hydroxybutyrate with a high molecular weight by methanotroph from methane and methanol. J Nat Gas Chem 17:103–109

    Article  Google Scholar 

Download references

Acknowledgments

We thank Dr. Lisa Stein and the Carbon-one Oxidation Network for the phaC gene sequence for M. trichosporium OB3b. We also thank Holly Sewell for her help in the laboratory.

This work was supported by a Graduate Research Fellowship from the National Science Foundation to AJP and by the California Environmental Protection Agency, Department of Toxic Substances Control under contract 07T3451.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Craig S. Criddle.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pieja, A.J., Rostkowski, K.H. & Criddle, C.S. Distribution and Selection of Poly-3-Hydroxybutyrate Production Capacity in Methanotrophic Proteobacteria. Microb Ecol 62, 564–573 (2011). https://doi.org/10.1007/s00248-011-9873-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00248-011-9873-0

Keywords

Navigation