Skip to main content
Log in

A Pseudomonas strain accumulating polyesters of 3-hydroxybutyric acid and medium-chain-length 3-hydroxyalkanoic acids

  • Biotechnology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Summary

A citronellol-utilizing bacterium was isolated that accumulated a polyester consisting of 3-hydroxybutyric acid (3HB) and of medium-chain-length 3-hydroxyalkanoic acids (3HAMCL) from various carbon sources up to approximately 70% of the cellular dry matter if the cells were cultivated in ammineral salts medium under nitrogen limitation. In octanoate-grown cells, for instance, the polyester consisted of 87.5 mol% 3HB and 12.5 mol% 3-hydroxyoctanoic acid (3HO), whereas it consisted of 10.3 mol% 3HB, 16.7 mol% 3HO and 73.0 mol% 3-hydroxydecanoic acid (3HD) in gluconate-grown cells. However, the results of various experiments indicated that a blend rather than a copolyester was synthesized in the cell. It was the only strain among 45 different recently isolated citronellol-utilizing bacteria that accumulated such a polyester. All other citronellol-utilizing bacteria behaved like Pseudomonas aeruginosa with respect to their polyhydroxyalkanoic acid (PHA) biosynthetic capabilities and accumulated PHA consisting of 3HAMCL with 3HO and 3HD as the main constituents from octanoate or gluconate, respectively, whereas 3HB was never present. None of 232 different heavy-metal-resistant bacteria was able to accumulate PHA composed of 3HB plus, for example, 3HO. Only 20.3% did not accumulate any PHA at all, 44.8% accumulated PHB from gluconate, and 34.9% behaved like P. aeruginosa. Many bacteria belonging to the latter group were distinguished from the other by rapid growth in nutrient broth and in gluconate mineral salts medium and by their ability to grow in the presence of a high concentration (up to 1.5%, w/v) of octanoate.

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

  • Ajello GW, Hoadley AW (1976) Fluorescent pseudomonads capable of growth at 41°C but distinct from Pseudomonas aeruginosa. J Clin Microbiol 4: 443–449

    Google Scholar 

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

    CAS  PubMed  Google Scholar 

  • Brandl H, Gross RA, Lenz RW, Fuller RC (1988) Pseudomonas oleovorans as a source of poly(β-hydroxyalkanoates) for potential applications as biodegradable polyesters. Appl Environ Microbiol 54: 1977–1982

    Google Scholar 

  • Brandl H, Gross RA, Knee EJ, Lenz RW, Fuller RC (1989) The ability of the phototrophic bacterium Rhodospirillum rubrum to produce various poly(β-hydroxyalkanoates): potential sources for biodegradable polyesters. Int J Biol Macromol 11: 49–56

    Google Scholar 

  • Doi Y, Tamaki A, Kunioka M, Soga K (1987) Biosynthesis of terpolyesters of 3-hydroxybutyrate, 3-hydroxyvalerate, and 5-hydroxyvalerate in Alcaligenes eutrophus from 5-chloropentanoic and pentanoic acids. Makromol Chem Rapid Commun 8: 631–635

    Google Scholar 

  • Fonseca-Wollheim F da, Bergmeyer HU, Gutamann I (1974) Ammoniak. In: Bergmeyer HU (ed) Methoden der enzymatischen Analyse, 3rd edn, Verlag Chemie, Weinheim, pp 1850–1853

    Google Scholar 

  • Haywood GW, Anderson AJ, Dawes EA (1989a) The importance of PHB-synthase substrate specificity in polyhydroxyalkanoate synthesis by Alcaligenes eutrophus. FEMS Microbiol Lett 57: 1–6

    Google Scholar 

  • Haywood GW, Anderson AJ, Dawes EA (1989b) A survey of the accumulation of novel polyhydroxyalkanoates by bacteria. Biotechnol Lett 11: 471–476

    Google Scholar 

  • Haywood GW, Anderson AJ, Ewing DF, Dawes EA (1990) Accumulation of a polyhydroxyalkanoate containing primarily 3-hydroxydecanoate from simple carbohydrate substrates by Pseudomonas sp. strain NCIMB 40135. Appl Environ Microbiol 56: 3354–3359

    Google Scholar 

  • Haywood GW, Anderson AJ, Williams DR, Dawes EA, Ewing DF (1991) The accumulation of a polyhydroxyalkanoate copolymer containing primarily 3-hydroxyvalerate from simple carbohydrate substrates by Rhodococcus sp. NCIMB 40126. Int J Biol Macromol 13: 83–88

    Google Scholar 

  • Huisman GW, Leeuw O de, Eggink G, Witholt B (1989) Synthesis of poly-3-hydroxyalkanoates is a common feature of fluorescent pseudomonads. Appl Environ Microbiol 55: 1949–1954

    Google Scholar 

  • Huisman GW, Wonink E, Meima R, Kazemier B, Terpstra P, Witholt B (1991) Metabolism of poly(3-hydroxyalkanoates) by Pseudomonas oleovorans: identification and sequences of genes and function of the encoded proteins in the synthesis and degradation of PHA. J Biol Chem 266: 2191–2198

    Google Scholar 

  • Kado CI, Liu ST (1981) Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol 145: 757–767

    Google Scholar 

  • Kunioka M, Nakamura Y, Doi Y (1988) New bacterial copolyester produced in Alcaligenes eutrophus from organic acids. Polym Commun 29: 174–176

    Google Scholar 

  • Lageveen RG, Huisman GW, Preusting H, Ketelaar P, Eggink G, Witholt B (1988) Formation of polyesters by Pseudomonas oleovorans: effect of substrates on formation and composition of poly-(R)-3-hydroxyalkanoates and poly-(R)-3-hydroxyalkenoates. Appl Environ Microbiol 54: 2924–2932

    Google Scholar 

  • Lenz RW, Kim BW, Ulmer HW, Fritzsche K, Knee E, Fuller RC (1990) Functionalized poly-β-hydroxyalkanoates produced by bacteria. In: Dawes EA (ed) Novel biodegradable microbial polymers. Kluwer Academic Publishers, Dordrecht, pp 23–35

    Google Scholar 

  • Liebergesell M, Hustede E, Timm A, Steinbüchel A, Fuller RC, Lenz RW, Schlegel HG (1991) Formation of poly(3-hydroxyalkanoates) by phototrophic and chemolithotrophic bacteria. Arch Microbiol 155: 415–421

    Google Scholar 

  • Lowry OH, Rosebrough HJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275

    CAS  PubMed  Google Scholar 

  • Preusting H, Kingma J, Huisman G, Steinbüchel A, Witholt B (1992) Formation of polyester blends by a recombinant strain of Pseudomonas oleovorans: different poly(3-hydroxyalkanoates) are stored in separate granules. J Environ Polym Degrad, in press

  • Ramsay BA, Saracovan I, Ramsay JA, Marchessault RH (1992) Effect of nitrogen limitation on long-side-chain poly-β-hydroxyalkanoate synthesis by Pseudomonas resinovorans. Appl Microbiol Biotechnol 58: 744–746

    Google Scholar 

  • Schlegel HG, Kaltwasser H, Gottschalk G (1961) Ein Submersverfahren zur Kultur wasserstoffoxidierender Bakterien: Wachstumsphysiologische Untersuchungen. Arch Mikrobiol 38: 209–222

    CAS  PubMed  Google Scholar 

  • Schmidt T, Schlegel HG (1989) Nickel and cobalt resistance of various bacteria isolated from soil and highly polluted domestic and industrial wastes. FEMS Microbiol Lett 62: 315–328

    Google Scholar 

  • Schubert P, Steinbüchel A, Schlegel HG (1988) Cloning of the Alcaligenes eutrophus genes for synthesis of poly-β-hydroxybutyric acid (PHB) and synthesis of PHB in Escherichia coli. J Bacteriol 170: 5837–5847

    Google Scholar 

  • Schubert P, Krüger N, Steinbüchel A (1991) Molecular analysis of the Alcaligenes eutrophus poly(3-hydroxybutyrate) biosynthetic operon: identification of the N terminus of poly(3-hydroxybutyrate) synthase and identification of the promoter. J Bacteriol 173: 168–175

    Google Scholar 

  • Seubert W (1960) Degradation of isoprenoid compounds by microorganisms. I. Isolation and characterization of an isoprenoid-degrading bacterium, Pseudomonas citronellois n. sp. J Bacteriol 79: 426–434

    Google Scholar 

  • Spurr AR (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26: 31–43

    CAS  PubMed  Google Scholar 

  • Steinbüchel A (1991a) Polyhydroxyalkanoic acids. In: Byrom D (ed) Biomaterials. MacMillan, London, pp 123–213

    Google Scholar 

  • Steinbüchel A (1991b) Polyhydroxyfettsäuren — thermoplastisch verformbare Polyester aus Bakterien. Nachrichten Chem Tech Lab 39: 1112–1124

    Google Scholar 

  • Steinbüchel A, Pieper U (1991) Production of a copolyester of 3-hydroxybutyric acid and 3-hydroxyvaleric acid from single unrelated carbon sources by a mutant of Alcaligenes eutrophus. Appl Microbiol Biotechnol 57: 1–7

    Google Scholar 

  • Steinbüchel A, Schlegel HG (1991) Physiology and molecular genetics of poly(β-hydroxyalkanoic acid) synthesis in Alcaligenes eutrophus. Mol Microbiol 5: 535–542

    Google Scholar 

  • Steinbüchel A, Schubert P (1989) Expression of the Alcaligenes eutrophus poly(β-hydroxybutyric acid)-synthetic pathway in Pseudomonas sp. Arch Microbiol 153: 101–104

    Google Scholar 

  • Timm A, Steinbüchel A (1990) Formation of polyesters consisting of medium-chain-length 3-hydroxyalkanoic acids from gluconate by Pseudomonas aeruginosa and other fluorescent pseudomonads. Appl Environ Microbiol 56: 3360–3367

    Google Scholar 

  • Timm A, Byrom D, Steinbüchel A (1990) Formation of blends of various poly(3-hydroxyalkanoic acids) by a recombinant strain of Pseudomonas oleovorans. Appl Microbiol Biotechnol 33: 296–301

    Google Scholar 

  • Timotius K, Schlegel HG (1987) Aus Abwässern isolierte nickelresistente Bakterien. Nachrichen der Akademie der Wissenschaften in Göttingen; II. Math.-Physik. Klasse, no. 3, pp 15–23

  • Valentin H, Schönebaum A, Steinbüchel A (1992) Identification of 4-hydroxyvaleric acid as a constituent in biosynthetic polyhydroxyalkanoic acids from bacteria. Appl Microbiol Biotechnol 36: 507–514

    Google Scholar 

  • Walther-Mauruschat A, Aragno M, Mayer F, Schlegel HG (1977) Micromorphology of Gram-negative hydrogen bacteria. II. Cell envelope, membranes, and cytoplasmic inclusions. Arch Microbiol 114: 101–110

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Correspondence to: A. Steinbüchel

Rights and permissions

Reprints and permissions

About this article

Cite this article

Steinbüchel, A., Wiese, S. A Pseudomonas strain accumulating polyesters of 3-hydroxybutyric acid and medium-chain-length 3-hydroxyalkanoic acids. Appl Microbiol Biotechnol 37, 691–697 (1992). https://doi.org/10.1007/BF00174829

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00174829

Keywords

Navigation