Skip to main content
SpringerLink
Log in

Studies on the Microbial Synthesis and Characterization of Polyhydroxyalkanoates Containing 4-Hydroxyvalerate Using γ-Valerolactone

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

Abstract

In this study, the ability of Cupriavidus sp. USMAA2-4 to synthesize polyhydroxyalkanoates (PHA) containing 4-hydroxyvalerate monomer (4HV) was studied through one-stage cultivation using γ-valerolactone as the carbon precursor. The presence of 4HV monomer unit in the polymer was detected through gas chromatography analysis, proving the capability of this wild strain bacterium to produce poly(3-hydrxybutyrate-co-3-hydroxyvalerate-co-4-hydroxyvalerate) [P(3HB-co-3HV-co-4HV)] terpolymer. Existence of a 4HV monomer unit in the PHA produced was further confirmed through 13C and 1H NMR analysis. P(3HB-co-88 % 3HV-co-1 % 4HV) terpolymer with the highest PHA content of 63 wt% was obtained through combination of 0.14 wt% C of γ-valerolactone with 0.42 wt% C of oleic acid. Various compositions of P(3HB-co-3HV-co-4HV) terpolymer with 3HV and 4HV compositions ranging from 11 to 94 mol% and from 1 to 4 mol%, respectively, were acquired by manipulating γ-valerolactone and oleic acid concentrations. The molecular weight and the thermal and mechanical properties of four different compositions of terpolymers—P(3HB-co-91 % 3HV-co-1 % 4HV), P(3HB-co-55 % 3HV-co-2 % 4HV), P(3HB-co-27 % 3HV-co-2 % 4HV), and P(3HB-co-9 % 3HV-co-1 % 4HV)—were characterized. Among these terpolymers, P(3HB-co-27 % 3HV-co-2 % 4HV) terpolymer with a molecular weight of 5.7 (105 Da) exhibited the highest elongation to break (264 %). The monomer unit compositional distributions of these terpolymers were investigated through acetone–water fractionation analysis. The results suggested that these produced terpolymers had broad 3HV compositional distribution and narrow 4HV compositional distribution.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Anderson, A. J., & Dawes, E. A. (1990). Microbiology Reviews, 54(4), 450–472.

    CAS  Google Scholar 

  2. Loo, C. Y., & Sudesh, K. (2007). Biosynthesis and native granule characteristics of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in Delftia acidovorans. International Journal of Biological Macromolecules, 40, 466–471.

    Article  CAS  Google Scholar 

  3. Steinbüchel, A., & Lutke-Eversloh, T. (2003). Metabolic engineering and pathway construction for biotechnological production of relevant polyhydroxyalkanoates in microorganisms. Biochemical Engineering Journal, 16, 81–96.

    Article  Google Scholar 

  4. Sudesh, K., Abe, H., & Doi, Y. (2000). Synthesis, structure and properties of polyhydroxyalkanoates: biological polyesters. Progress in Polymer Science, 25, 1503–1555.

    Article  CAS  Google Scholar 

  5. Amirul, A. A. A., Syairah, S. N., Yahya, A. R. M., Azizan, M. N. M., & Majid, M. I. A. (2008). Synthesis of biodegradable polyesters by Gram negative bacterium isolated from Malaysian environment. World Journal of Microbiology and Biotechnology, 24, 1327–1332.

    Article  CAS  Google Scholar 

  6. Schmack, G., Gorenflo, V., & Steinbüchel, A. (1998). Biotechnological production and characterization of polyesters containing 4-hydroxyvaleric acid and medium-chain-length hydroxyalkanoic acids. Macromolecules, 31(3), 644–649.

    Article  CAS  Google Scholar 

  7. Noda, I., Green, P. R., Satkowski, M. M., & Schechtman, L. A. (2005). Preparation and properties of a novel class polyhydroxyalkanoates copolymers. Biomacromolecules, 6, 580–586.

    Article  CAS  Google Scholar 

  8. Suriyamongkol, P., Weselake, R., Narine, S., Moloney, M., & Shah, S. (2007). Biotechnological approaches for the production of polyhydroxyalkanoates in microorganisms and plants—a review. Biotechnology Advances, 25(2), 148–175.

    Article  CAS  Google Scholar 

  9. Gorenflo, V., Schmack, G., Vogel, R., & Steinbüchel, A. (2001). Development of a process for the biotechnological large-scale production of 4-hydroxyvalerate-containing polyesters and characterization of their physical and mechanical properties. Biomacromolecules, 2, 45–57.

    Article  CAS  Google Scholar 

  10. Martin, C. H., & Prather, K. L. J. (2009). High-titer production of monomeric hydroxyvalerates from levulinic acid in Pseudomonas putida. Journal of Biotechnology, 139(1), 61–67.

    Article  CAS  Google Scholar 

  11. Yu, J., Chen, L. X. L., & Sato, S. (2009). Biopolyester synthesis and protein regulations in Ralstonia eutropha on levulinic acid and its derivatives from biomass refining. Journal of Biobased Materials and Bioenergy, 3(10), 113–112.

    Article  CAS  Google Scholar 

  12. Valentin, H. E., & Steinbüchel, A. (1995). Accumulation of poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid-co-4-hydroxyvaleric acid) by mutants and recombinant strains of Alcaligenes eutrophus. Journal of Polymers and the Environment, 3(3), 169–175.

    Article  CAS  Google Scholar 

  13. Ceisielski, S., Kwiatkowska, A. C., Pokoj, T., & Klimiuk, E. (2006). Molecular detection and diversity of medium-chain-length polyhydroxyalkanoates-producing bacteria enriched in activated sludge. Journal of Applied Microbiology, 101, 190–199.

    Article  Google Scholar 

  14. Vandamme, P., & Coenye, T. (2004). Taxonomy of the genus Cupriavidus: a tale of lost and found. International Journal of Systematic and Evolutionary Microbiology, 54, 2285–2289.

    Article  Google Scholar 

  15. Nakamura, S., Doi, Y., & Scandola, M. (1992). Microbial synthesis and characterization of poly(3-hydroxybutyrate-co-4-hydroxybutyrate). American Chemical Society, 25, 4237–4241.

    CAS  Google Scholar 

  16. Hiramitsu, M., Koyama, N., & Doi, Y. (1993). Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Alcaligenes latus. Biotechnology Letters, 15, 461–464.

    Article  CAS  Google Scholar 

  17. Lee, W. H., Loo, C. Y., Nomura, C. T., & Sudesh, K. (2008). Biosynthesis of polyhydroxyalkanoate copolymers from mixtures of plant oils and 3-hydroxyvalerate precursors. Bioresource Technology, 99(15), 6844–6851.

    Article  CAS  Google Scholar 

  18. Kahar, P., Tsuge, T., Taguchhi, K., & Doi, Y. (2004). High yield production of polyhydroxyalkanoates from soybean oil by Ralstonia eutropha and its recombinant strain. Polymer Degradation and Stability, 83, 79–86.

    Article  CAS  Google Scholar 

  19. Kunioka, M., Nakamura, S., & Doi, Y. (1988). Polymer Communication, 29, 174–176.

    CAS  Google Scholar 

  20. Ramachandran, H., Iqbal, N. M., Sipaut, C. S., & Abdullah, A. A. (2011). Biosynthesis and characterization of poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) terpolymer with various monomer compositions by Cupriavidus sp. USMAA2-4. Applied Biochemistry and Biotechnology, 164, 867–877.

    Google Scholar 

  21. Braunegg, G., Sonnleitner, B., & Lafferty, R. M. (1978). A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass. European Journal of Applied Microbiology and Biotechnology, 6(1), 29–37.

    Article  CAS  Google Scholar 

  22. Amirul, A., Yahya, A. R. M., Sudesh, K., Azizan, M. N. M., & Majid, M. I. A. (2009). Isolation of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) producer from Malaysian environment using γ-butyrolactone as carbon source. World Journal of Microbiology and Biotechnology, 25(7), 1199–1206.

    Article  CAS  Google Scholar 

  23. Vigneswari, S., Nik, L., Majid, M. I. A., & Amirul, A. A. (2009). Improved production of poly(3-hydroxybutyrate-co-4-hydroxbutyrate) copolymer using a combination of 1,4-butanediol and γ-butyrolactone. World Journal of Microbiology and Biotechnology, 26(4), 743–746.

    Article  Google Scholar 

  24. Mitomo, H., Morishita, N., & Doi, Y. (1995). Structural changes of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) fractionated with acetone–water solution. Polymer, 36(13), 2573–2578.

    Article  CAS  Google Scholar 

  25. Valentin, H. E., Schönebaum, A., & Steinbüchel, A. (1992). Identification of 4-hydroxyvaleric acid as a constituent of biosynthetic polyhydroxyalkanoic acids from bacteria. Applied Microbiology and Biotechnology, 36(4), 507–514.

    Article  CAS  Google Scholar 

  26. Lee, E. Y., & Choi, C. Y. (1997). Structural identification of polyhydroxyalkanoic acid (PHA) containing 4-hydroxyalkanoic acids by gas chromatography–mass spectrometry (GC-MS) and its application to bacteria screening. Biotechnology Techniques, 11(3), 167–171.

    Article  CAS  Google Scholar 

  27. Bloembergen, S., Holden, D. A., Hamer, G. K., Bluhm, T. L., & Marchessault, R. H. (1986). Studies of composition and crystallinity of bacterial poly(β-hydroxybutyrate-co-β-hydroxyvalerate). Macromolecules, 19, 2865–2871.

    Article  CAS  Google Scholar 

  28. Amirul, A. A., Yahya, A. R. M., Sudesh, K., Azizan, M. N. M., & Majid, M. I. A. (2008). Biosynthesis of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer by Cupriavidus sp. USMAA1020 isolated from Lake Kulim, Malaysia. Bioresource Technology, 99(11), 4903–4909.

    Article  CAS  Google Scholar 

  29. Rahayu, A., Zaleha, Z., Yahya, A. R. M., Majid, M. I. A., & Amirul, A. A. (2008). Production of copolymer poly(3-hydroxybutyrate-co-4-hydroxybutyrate) through a one-step cultivation process. World Journal of Microbiology and Biotechnology, 24(11), 2403–2409.

    Article  CAS  Google Scholar 

  30. Slater, S., Houmiel, K. L., Tran, M., Mitsky, T. A., Taylor, N. B., Padgette, S. R., & Gruys, K. J. (1998). Multiple ß-ketothiolases mediate poly(ß-hydroxyalkanoate) copolymer synthesis in Ralstonia eutropha. Journal of Bacteriology, 180, 1979–1987.

    CAS  Google Scholar 

  31. Lo, K., Chua, H., Lawford, H., Lo, W., & Yu, P. (2005). Effects of fatty acids on growth and poly-3-hydroxybutyrate production in bacteria. Applied Biochemistry and Biotechnology, 122, 575–580.

    Article  Google Scholar 

  32. Lee, S. Y., Lee, Y. K., & Chang, H. N. (1995). Stimulatory effects of amino acids and oleic acid on poly(3-hydroxybutyric acid) synthesis by recombinant Escherichia coli. Journal of Fermentation and Bioengineering, 79(2), 177–180.

    Article  CAS  Google Scholar 

  33. Chanprateep, S., & Kulpreecha, S. (2006). Production and characterization of biodegradable terpolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) by Alcaligenes sp. A-04. Journal of Bioscience and Bioengineering, 101, 51–56.

    Article  CAS  Google Scholar 

  34. Ng, K. S., Wong, Y. M., Tsuge, T., & Sudesh, K. (2011). Biosynthesis and characterization of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) copolymers using jatropha oil as the main carbon source. Process Biochemistry, 46, 1572–1578.

    Article  CAS  Google Scholar 

  35. Luo, S., Grubb, D. T., & Netravali, A. N. (2002). The effect of molecular weight on the lamellar structure, thermal and mechanical properties of poly(hydroxybutyrate-co-hydroxyvalerate). Polymer, 43, 4159–4166.

    Article  CAS  Google Scholar 

  36. Rehm, B. H. A. (2003). Polyester synthases: natural catalysts for plastics. Biochemistry Journal, 376, 15–33.

    Article  CAS  Google Scholar 

  37. Riande, E., Diaz-Calleja, R., Prolongo, M. G., Masegosa, R. M., & Salom, C. (2000). Polymer viscoelasticity, stress and strain in practice. New York: Marcel Dekker.

    Google Scholar 

  38. Scandola, M., Ceccorulli, G., Pizzoli, M., & Gazzano, M. (1992). Study of the crystal phase and crystallization rate of bacterial poly(3-hydroxybutyrate-co-3-hydroxyvalerate). Macromolecules, 25, 1405–1410.

    Article  CAS  Google Scholar 

  39. Savenkova, L., & Gercberga, Z. (2000). Effect of 3-hydroxyvalerate content on some physical and mechanical properties of polyhydroxyalkanoates produced by Azotobacter chroococcum. Process Biochemistry, 36(5), 445–450.

    Article  CAS  Google Scholar 

  40. Fahima Azira, T., Nursolehah, A. A., Norhayati, Y., Majid, M. I. A., & Amirul, A. A. (2011). Biosynthesis of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) terpolymer by Cupriavidus sp. USMAA2-4 through two-step cultivation process. World Journal of Microbiology and Biotechnology, 1–9.

  41. Doi, Y. (1990). Microbial Polyesters. New York: VCH Publishers, Inc.

    Google Scholar 

  42. Barham, P. J., & Keller, A. (1986). The relationship between microstructure and mode of fracture in polyhydroxybutyrate. Journal of Polymer Science Polymer Physics Edition, 24, 69–77.

    Article  CAS  Google Scholar 

  43. Zhao, W., & Chen, G. Q. (2007). Production and characterization of terpolyester poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) by recombinant Aeromonas hydrophila 4AK4 harboring genes phaAB. Process Biochemistry, 42, 1342–1347.

    Article  CAS  Google Scholar 

  44. Yamada, S., Wang, Y., Asakawa, N., Yoshie, N., & Inoue, Y. (2001). Crystalline structural change of bacterial poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with narrow compositional distribution. Macromolecules, 34, 4659–4661.

    Article  CAS  Google Scholar 

  45. Cao, A., Ichikawa, M., Kasuya, K. I., Yoshie, N., Asakawa, N., Inoue, Y., Doi, Y., & Abe, H. (1996). Composition fractionation and thermal characterization of poly(3-hydroxybutyrate-co-3-hydroxypropionate). Polymer Journal, 28(12), 1096–1102.

    Article  CAS  Google Scholar 

  46. Feng, L., Watanabe, T., Wang, Y., Kichise, T., Fukuchi, T., Chen, G. Q., Doi, Y., & Inoue, Y. (2002). Studies on comonomer compositional distribution of bacterial poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)s and thermal characteristics of their fractions. Biomacromolecules, 3, 1071–1077.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Biological Sciences, Universiti Sains Malaysia, 11800, Penang, Malaysia

    A. R. Muzaiyanah & A. A. Amirul

  2. Malaysian Institute of Pharmaceutical and Nutraceuticals, MOSTI, Penang, Malaysia

    A. A. Amirul

Authors
  1. A. R. Muzaiyanah
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. A. Amirul
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A. A. Amirul.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Muzaiyanah, A.R., Amirul, A.A. Studies on the Microbial Synthesis and Characterization of Polyhydroxyalkanoates Containing 4-Hydroxyvalerate Using γ-Valerolactone. Appl Biochem Biotechnol 170, 1194–1215 (2013). https://doi.org/10.1007/s12010-013-0247-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-013-0247-6

Keywords

Search

Navigation