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Biosynthetic enhancement of single-stage Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) production by manipulating the substrate mixtures

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Journal of Industrial Microbiology & Biotechnology

Abstract

Two-stage fermentation was normally employed to achieve a high poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] productivity with higher 4HB molar fraction. Here, we demonstrated single-stage fermentation method which is more industrial feasible by implementing mixed-substrate cultivation strategy. Studies on bioreactor scale show a remarkably high PHA accumulation of 73 wt%, contributing to a high PHA concentration and product yield of 8.6 g/L and 2.7 g/g, respectively. This fermentation strategy has resulted in copolymers with wider range of 4HB monomer composition, which ranges from 12 to 55 mol%. These copolymers show a broad range of weight average molecular weight (M w ) from 119.5 to 407.0 kDa. The copolymer characteristics were found to be predominantly affected by the nature of the substrates and the mixture strategies, regardless of the 4HB monomer compositions. This was supported by the determination of copolymer randomness using 13C-NMR analysis. The study warrants significantly in the copolymer scale-up and modeling at industrial level.

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References

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

    Article  CAS  Google Scholar 

  2. Anderson AJ, Dawes EA (1999) Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev 54(4):450–472

    Google Scholar 

  3. Babel W, Brinkmann U, Muller RH (1993) The auxiliary substrate concept—an approach for overcoming limits of microbial performances. Acta Biotechnol 13:211–242

    Article  CAS  Google Scholar 

  4. Braunegg G, Sonnleitner B, Lafferty RM (1978) A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass. Appl Microbiol Biotechnol 6:29–37

    Article  CAS  Google Scholar 

  5. Cavalheiro JMBT, Raposo RS, de Almeida MCMD, Cesario MT, Sevrin C, Grandfils C, da Fonseca MMR (2012) Effect of cultivation parameters on the production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) and poly(3-hydroxybutyrate-4-hydroxybutyrate-3-hydroxyvalerate) by Cupriavidus necator using waste glycerol. Biores Technol 111:391–397

    Article  CAS  Google Scholar 

  6. Cesario MT, Raposo RS, de Almeida MCMD, van Keulen F, Ferreira BS, Telo JP, de Fonseca MMR (2014) Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Burkholderia sacchari using wheat straw hydrolysates and gamma-butyrolactone. Int J Bio Macromol 71:59–67

    Article  CAS  Google Scholar 

  7. Chai H-L, Ahmad R, Yahya ARM, Majid MIA, Amirul AA (2009) Microbial synthesis of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer by Cupriavidus sp. USMAA2-4 through a two-step cultivation process. African. J Biotechnol 8(17):4189–4196

    CAS  Google Scholar 

  8. Choi MH, Yoon SC, Lenz RW (1999) Production of Poly(3-Hydroxybutyric acid-co-4-Hydroxybutyric acid) and Poly(4-Hydroxybutyric acid) without subsequent degradation by Hydrogenophaga pseudoflava. Appl Environ Microbiol 65(4):1570–1577

    PubMed Central  CAS  PubMed  Google Scholar 

  9. Gumel AM, Annuar MSM, Heidelberg T (2012) Effects of carbon substrates on biodegradable polymer composition and stability produced by Delftia tsuruhatensis Bet002 isolated from palm oil mill effluent. Polym Degrad Stab 97(8):1224–1231

    Article  CAS  Google Scholar 

  10. Harder W, Dijkhuizen L (1982) Strategies of Mixed Substrate Utilization in Microorganisms. Phil Trans R Soc Lond B 297:459–480

    Article  CAS  Google Scholar 

  11. Hiramitsu M, Koyama N, Doi Y (1993) Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Alcaligenes latus. Biotechnol Lett 15(5):461–464

    Article  CAS  Google Scholar 

  12. Huong K-H, Yahya ARM, Amirul AA (2014) Pronounced synergistic influence of mixed substrate cultivation on single step copolymer P(3HB-co-4HB) biosynthesis with a wide range of 4HB monomer composition. J Chem Technol Biotechnol 89(7):1023–1029

    Article  CAS  Google Scholar 

  13. Kamiya N, Yamamoto Y, Inoue Y, Chujo R, Doi Y (1989) Microstructure of bacterially synthesized poly(3-hydroxybutyrate-co-3-hydroxyvalerate). Macromol 22:1676–1682

    Article  CAS  Google Scholar 

  14. Kang C-K, Kusaka S, Doi Y (1995) Structure and properties of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) produced by Alcaligenes latus. Biotechnol Lett 17(6):583–588

    Article  CAS  Google Scholar 

  15. Kim JS, Lee BH, Kin BS (2005) Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Ralstonia eutropha. Biochem Eng J 23:169–174

    Article  CAS  Google Scholar 

  16. Kimura H, Iwama M, Sasaki S, Takeishi M (1999) Production of polyester blend of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) and poly(3-hydroxyalkanoate) with saturated and unsaturated monomers from 4-hydroxybutyric acid by Chromobacterim sp. Chem Lett 28:737–738

    Article  Google Scholar 

  17. Lee W-H, Azizan NM, 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 

  18. Loo C-Y, Sudesh K (2007) Polyhydroxyalkanoates: bio-based microbial plastic and their properties. M Polym J 2:31–57

    Google Scholar 

  19. Majid MIA, Ismail J, Few LL, Tan CF (2002) The degradation kinetics of poly(3-hydroxybutyrate) under non-aqueous and aqueous conditions. Eur Polym J 38:837–839

    Article  CAS  Google Scholar 

  20. Martin DP, Williams SF (2003) Medical applications of poly-4-hydroxybutyrate: a strong flexible absorbable biomaterial. Biochem Eng J 16:97–105

    Article  CAS  Google Scholar 

  21. Mitomo H, Hsieh WC, Nishiwaki K, Kasuya K, Doi Y (2001) Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) produced by Comamonas acidovarans. Polym 42:3455–3461

    Article  CAS  Google Scholar 

  22. Ramachandran H, Amirul AA (2013) Bioconversion of glycerine pitch into a novel yellow-pigmented P(3HB-co-4HB) copolymer: synergistic effect of ammonium acetate and polymer characteristics. Appl Biochem Biotechnol 172(2):891–909

    Article  Google Scholar 

  23. Ramachandran H, Amirul AA (2013) Yellow-pigmented Cupriavidus sp., a novel bacterium capable of utilizing glycerine pitch for the sustainable production of P(3HB-co-4HB). J Chem Technol Biotechnol 88(6):1030–1038

    Article  CAS  Google Scholar 

  24. Rao U, Sridhar R, Sehgal PK (2010) Biosynthesis and biocompatibility of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) produced by Cupriavidus necator from spent palm oil. Biochem Eng J 49:13–20

    Article  CAS  Google Scholar 

  25. Renner G, Pongratz K, Braunegg G (1996) production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Comamonas testoteronii A3. Food Technol Biotechnol 34:91–95

    CAS  Google Scholar 

  26. 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 

  27. Vigneswari S, Nik LA, Majid MIA, Amirul AA (2010) Improved production of poly(3-hydroxybutyrate-co-4-hydroxbutyrate) copolymer using a combination of 1,4-butanediol and γ-butyrolactone. World J Microbiol Biotechnol 26:743–746

    Article  CAS  Google Scholar 

  28. Vigneswari S, Vijaya S, Majid MA, Sudesh K, Sipaut CS, Azizan MNM, Amirul AA (2009) Enhanced production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer with manipulated variables and its properties. J Ind Microbiol Biotechnol 36:547–556

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The authors acknowledge the research grants provided by Ministry of Science, Technology and Innovation (02-05-23-SF0003) and also Malaysia Toray Science Foundation (MTSF) that has resulted in this article. We are also grateful to Mr. Leonard Lu Tze Jian for his help in language editing.

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Authors and Affiliations

  1. Malaysian Institute of Pharmaceuticals and Nutraceuticals, NIBM, 11700, Gelugor, Penang, Malaysia

    Kai-Hee Huong & A. A. Amirul

  2. School of Biological Sciences, Universiti Sains Malaysia, 11800, Minden, Penang, Malaysia

    Shantini Kannusamy & A. A. Amirul

  3. School of Chemical Sciences, Universiti Sains Malaysia, 11800, Minden, Penang, Malaysia

    Sumithda Yeong Hui Lim

  4. Centre for Chemical Biology, Universiti Sains Malaysia, 11900, Bayan Lepas, Penang, Malaysia

    A. A. Amirul

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  1. Kai-Hee Huong
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  2. Shantini Kannusamy
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  3. Sumithda Yeong Hui Lim
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  4. A. A. Amirul
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Correspondence to A. A. Amirul.

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Huong, KH., Kannusamy, S., Lim, S.Y.H. et al. Biosynthetic enhancement of single-stage Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) production by manipulating the substrate mixtures. J Ind Microbiol Biotechnol 42, 1291–1297 (2015). https://doi.org/10.1007/s10295-015-1657-y

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  • DOI: https://doi.org/10.1007/s10295-015-1657-y

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