Skip to main content
SpringerLink
Log in

Biosynthesis of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) terpolymer by Cupriavidus sp. USMAA2-4 through two-step cultivation process

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

A locally isolated Gram-negative bacterium, Cupriavidus sp. USMAA2-4 was found capable of producing terpolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) [P(3HB-co-3HV-co-4HB)] using γ-butyrolactone or 1,4-butanediol with either valeric acid or 1-pentanol as the carbon source. The present of 3HB, 3HV and 4HB monomers were confirmed by gas chromatography (GC) and nuclear magnetic resonance (NMR) analysis. PHA concentration of 1.9 g/l was the highest value obtained using the combination of 1,4-butanediol and 1-pentanol through one-step cultivation process. PHA concentration obtained through two-step cultivation process was higher for all the combinations and the highest value achieved was 2.5 g/l using γ-butyrolactone and 1-pentanol as carbon source. Various molar fractions of 4HB and 3HV ranging from 6 to 14 mol% and 39 to 87 mol%, respectively were produced through two-step cultivation process by manipulating the concentration of γ-butyrolactone. As the culture aeration was reduced, the molar fraction of 3HV and 4HB increased from 40 to 67 mol% and 10 to 24 mol%, respectively while the dry cell weight and PHA content decreased. The terpolymer produced was characterized using gel permeation chromatography (GPC) and differential scanning calorimetry (DSC). The number-average molecular weight (M n) and the melting temperature (T m)) of the terpolymer were in the range of 177–484 kDa and 160–164°C, respectively.

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

Similar content being viewed by others

References

  • Amirul AA, Yahya ARM, Sudesh K, Azizan MNM, Majid MIA (2008a) Biosynthesis of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer by Cupriavidus sp. USMAA1020 isolated from Lake Kulim, Malaysia. Bioresour Technol 99:4903–4909

    Article  CAS  Google Scholar 

  • Amirul AA, Syairah SN, Yahya ARM, Azizan MNM, Majid MIA (2008b) Synthesis of biodegradable polyesters by Gram-negative bacterium from Malaysian environment. World J Microbiol Biotechnol 24(8):1327–1332

    Article  CAS  Google Scholar 

  • Anderson AJ, Dawes EA (1990) Occurrence, metabolic role and industrial use of bacteria polyhydroxylalkanoates. Microbiol Rev 54:450–472

    CAS  Google Scholar 

  • Braunegg G, Sonnleitner B, Lafferty RM (1978) A rapid 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 

  • Braunegg G, Lefebvre G, Genser KF (1998) Polyhydroxyalkanoates, biopolyesters from renewable resources: physiological and engineering aspects. J Biotechnol 65:127–161

    Article  CAS  Google Scholar 

  • Dawes EA, Senior PJ (1973) The role and regulation energy reserve polymers in microorganisms. In: Rose AH, Tempest DW (eds) Advances in microbial physiologically. Acad Press, London, p 140

    Google Scholar 

  • Doi Y, Kunioka M, Nakamura Y, Soga K (1986) Nuclear magnetic resonance studies on poly (β-hydroxybutyrate and a copolyester of β-hydroxybutyrate and β-hydroxyvalerate isolated from Alcaligenes eutrophus H16. Macromol 19:2860–2864

    Article  CAS  Google Scholar 

  • Fukui T, Kichise T, Yoshida Y, Doi Y (1997) Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyheptanoate) terpolymers by recombinant Alcaligenes eutropus. Biotechnol Lett 19(11):1093–1097

    Article  CAS  Google Scholar 

  • Grothe E, Young MM, Chisti Y (1999) Fermentation optimization for the production of poly(β-hydroxybutyric acid) microbial thermoplastic. Enzyme Microb Technol 25:132–141

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Kimura H, Yoshida Y, Doi Y (1992) Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Pseudomonas acidovorans. Biotechnol Lett 14:445

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Lee WH, Azizan MNM, Sudesh K (2004) Effects of culture conditions on the composition of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) synthesized by Comamonas acidovorans. Polym Degrad Stab 84:129–134

    Article  CAS  Google Scholar 

  • Lee WH, Loo CY, Nomura CI, Sudesh K (2008) Biosynthesis of polyhydroxyalkanaote copolymers from mixtures of plant oils and 3-hydroxyvalerate precursors. Bioresour Technol 99:6844–6851

    Article  CAS  Google Scholar 

  • Lemos PC, Serafirm LS, Reis MAM (2006) Synthesis of polyhydroxyalkanoates from different short-chain fatty acids by mixed cultures submitted to aerobic dynamic feeding. Biotechnol 122:226–238

    Article  CAS  Google Scholar 

  • Madden LA, Anderson AJ, Asrar J, Berger P, Garrett P (2000) Production and characterization of poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) synthesized by Ralstonia eutropha in fed-batch cultures. Polym 41:3499–3505

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Marchessault RH (1996) Tender morsels for bacteria: recent development in microbial polyesters. Trends Polym Sci 4:163–168

    CAS  Google Scholar 

  • Mukhopadhyay M, Patra A, Paul AK (2005) Production of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Rhodopseudomonas palustris SP5212. World J of Microbiol Biotechnol 21:765–769

    Article  CAS  Google Scholar 

  • Park SJ, Ahn WS, Green PR, Lee SY (2001) Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) by metabolically engineered Escherichia coli strains. Biotechnol Bioeng 74(1):81–86

    Article  CAS  Google Scholar 

  • Peoples OP, Sinskey AJ (1989) Poly-beta-hydroxybutyrate (PHB) biosynthesis in Alcaligenes eutrophus H16: identification and characterization of the PHB polymerase gene (phbC). J Biol Chem 264:15298–15303

    CAS  Google Scholar 

  • Rahayu A, Zaleha Z, Yahya ARM, Majid MIA, Amirul AA (2008) Production of copolymer poly(3-hydroxybutyrate-co-4-hydroxybutyrate) through a one-step cultivation process. World J Microbiol Biotechnol 24:2403–2409

    Article  CAS  Google Scholar 

  • Reddy CSK, Ghai R, Rashmi KVC (2003) Polyhydroxyalkanoate—an overview. Bioresour Technol 87:137–146

    Article  CAS  Google Scholar 

  • Saito Y, Doi Y (1994) Microbial synthesis and properties of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) in Comamonas acidovorans. Int Biol J Macromol 16:99–104

    Article  CAS  Google Scholar 

  • Saito Y, Nakamura S, Hiramitsu M, Doi Y (1996) Microbial synthesis and properties of poly(3-hydroxybutyrate-co-4-hydroxybutyrate). Polym Int 39:169–174

    Article  CAS  Google Scholar 

  • Steinbüchel A, Lutke-Eversloh T (2003) Metabolic engineering and pathway construction for biotechnological production of relevant polyhydroxyalkanoates in microorganims. Biochem 16:81–96

    Google Scholar 

  • Sudesh K, Abe H, Doi Y (2000) Synthesis, structure and properties of polyhydroxyalkanoates: biological polyester. Prog Polym Sci 25:1503–1555

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Valentin HE, Lee EY, Choi CY, Steinbüchel A (1994) Identififcation of 4-hydroxyhexanoic acids as a new constituent of biosynthetic polyhydroxyalkanoic acids from bacteria. Appl Microbiol Biotechnol 40:710–716

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors acknowledged the research grant provided by MOSTI, Malaysia (Science Fund, 02-01-05-SF0363) that has resulted in this article.

Author information

Authors and Affiliations

  1. School of Biological Sciences, Universiti Sains Malaysia, 11800, Pulau Pinang, Malaysia

    A. A. Nursolehah, Y. Norhayati & A. A. Amirul

  2. Malaysian Institute of Pharmaceutical and Nutraceutical, MOSTI, Malaysia

    T. M. Fahima Azira, M. I. A. Majid & A. A. Amirul

Authors
  1. T. M. Fahima Azira
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. A. Nursolehah
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y. Norhayati
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. I. A. Majid
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. 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

Fahima Azira, T.M., Nursolehah, A.A., Norhayati, Y. et al. Biosynthesis of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) terpolymer by Cupriavidus sp. USMAA2-4 through two-step cultivation process. World J Microbiol Biotechnol 27, 2287–2295 (2011). https://doi.org/10.1007/s11274-011-0693-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-011-0693-7

Keywords

Search

Navigation