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Journal of Polymer Research

, 24:158 | Cite as

Extraction and purification of Polyhydroxyalkanoates (PHAs): application of Thermoseparating aqueous two-phase extraction

  • Yoong Kit Leong
  • Pau Loke Show
  • John Chi-Wei Lan
  • Hwei-San Loh
  • Yee Jiun Yap
  • Tau Chaun Ling
ORIGINAL PAPER

Abstract

Being biodegradable, non-toxic and renewable as well as having similar or better properties than commercial plastics, polyhydroxyalkanoates (PHAs) can be a potential game changer in the polymer industry. Although viewed as a sustainable alternative to petrochemicals due to its biodegradability, PHAs are plagued with low commercial value due to their high production and recovery costs. Having the benefits of providing a mild environment for bioseparation, being environment-friendly and scalable, together with it its distinctive thermoseparating properties and ease of recyclability, thermoseparating-based aqueous two-phase extraction (ATPE) has provided the eco-friendly and economical solution to the PHA dilemma. ATPE-influencing factors such as types of thermoseparating polymer, concentration of phase-forming components, pH, and effect of centrifugation were investigated. Under the condition of 14 wt/wt% of EOPO 3900 concentration, 14 wt/wt% of ammonium sulfate concentration and pH 6 without the needs for extra centrifugation steps, a recovery yield and a purification factor of up to 72.2% and 1.61 fold can be achieved with the copolymers which can be recycled and reused twice. Thermoseparating ATPE has thus been proven to be a powerful primary purification tool for PHAs.

Keywords

Aqueous two-phase extraction Bioseparation Downstream processing Polyhydroxyalkanoates Purification Thermoseparating polymers 

Abbreviations

ATPE

Aqueous two-phase extraction

ATPS

Aqueous two-phase system

CP

Cloud point

EOPO

Ethylene oxide-propylene oxide

Kpa

Partition coefficient

LCST

Lower critical solution temperature

PEG

Polyethylene glycol

PF

Purifycation factor

TLL

Tie-line length

TSP

Thermoseparating polymer

Vr

Volume ratio

Y

Yield

Notes

Acknowledgements

This work is supported financially by Fundamental Research Grant Scheme (Malaysia, FRGS/1/2013/SG05/UNIM/02/1), MyBrain15 (MyPhD), Ministry of Science, Technology and Innovation (MOSTI-02-02-12-SF0256) and National Science Council (Taiwan, NSC102-2221-E-155-057 and NSC101-2632-E-155-001-MY3).

References

  1. 1.
    Castilho LR, Mitchell DA, Freire DM (2009) Production of polyhydroxyalkanoates (PHAs) from waste materials and by-products by submerged and solid-state fermentation. Bioresour Technol 100:5996–6009CrossRefGoogle Scholar
  2. 2.
    Leong YK, Show PL, Ooi CW, Ling TC, Lan JC (2014) Current trends in polyhydroxyalkanoates (PHAs) biosynthesis: insights from the recombinant Escherichia Coli. J Biotechnol 180:52–65CrossRefGoogle Scholar
  3. 3.
    Keshavarz T, Roy I (2010) Polyhydroxyalkanoates: bioplastics with a green agenda. Curr Opin Microbiol 13:321–326CrossRefGoogle Scholar
  4. 4.
    Gumel AM, Annuar MSM, Chisti Y (2013) Recent advances in the production, recovery and applications of Polyhydroxyalkanoates. J Polym Environ 21:580–605CrossRefGoogle Scholar
  5. 5.
    Jacquel N, Lo C-W, Wei Y-H, Wu H-S, Wang SS (2008) Isolation and purification of bacterial poly(3-hydroxyalkanoates). Biochem Eng J 39:15–27CrossRefGoogle Scholar
  6. 6.
    Kepka C, Rhodin J, Lemmens R, Tjerneld F, Gustavsson P-E (2004) Extraction of plasmid DNA from Escherichia coli cell lysate in a thermoseparating aqueous two-phase system. J Chrom A 1024:95–104CrossRefGoogle Scholar
  7. 7.
    Karkaş T, Önal S (2012) Characteristics of invertase partitioned in poly(ethylene glycol)/magnesium sulfate aqueous two-phase system. Biochem Eng J 60:142–150CrossRefGoogle Scholar
  8. 8.
    Barbosa JM, Souza RL, Fricks AT, Zanin GM, Soares CM, Lima AS (2011) Purification of lipase produced by a new source of bacillus in submerged fermentation using an aqueous two-phase system. J Chrom B, Analytical technologies in the biomedical and life sciences 879:3853–3858CrossRefGoogle Scholar
  9. 9.
    Li M, Peeples TL (2004) Purification of hyperthermophilic archaeal amylolytic enzyme (MJA1) using thermoseparating aqueous two-phase systems. J Chrom B, Analytical technologies in the biomedical and life sciences 807:69–74CrossRefGoogle Scholar
  10. 10.
    Divyashree MS, Shamala TR, Rastogi NK (2009) Isolation of polyhydroxyalkanoate from hydrolyzed cells of Bacillus flexus using aqueous two-phase system containing polyethylene glycol and phosphate. Biotechnol Bioprocess Eng 14:482–489CrossRefGoogle Scholar
  11. 11.
    Diamond AD, Hsu JT (1992) Aqueous two-phase systems for biomolecule separation. Adv Biochem Eng Biotechnol 47:89–135Google Scholar
  12. 12.
    Vazquez-Villegas P, Espitia-Saloma E, Rito-Palomares M, Aguilar O (2013) Low-abundant protein extraction from complex protein sample using a novel continuous aqueous two-phase systems device. J Sep Sci 36:391–399CrossRefGoogle Scholar
  13. 13.
    Dong B, Yuan X, Zhao Q, Feng Q, Liu B, Guo Y, Zhao B (2015) Ultrasound-assisted aqueous two-phase extraction of phenylethanoid glycosides from Cistanche deserticola Y. C Ma stems J Sep Sci 38:1194–1203CrossRefGoogle Scholar
  14. 14.
    Ng HS, Tan CP, Mokhtar MN, Ibrahim S, Ariff A, Ooi CW, Ling TC (2012) Recovery of Bacillus cereus cyclodextrin glycosyltransferase and recycling of phase components in an aqueous two-phase system using thermo-separating polymer. Sep Purif Technol 89:9–15CrossRefGoogle Scholar
  15. 15.
    Show PL, Tan CP, Shamsul Anuar M, Ariff A, Yusof YA, Chen SK, Ling TC (2012) Extractive fermentation for improved production and recovery of lipase derived from Burkholderia cepacia using a thermoseparating polymer in aqueous two-phase systems. Bioresour Technol 116:226–233CrossRefGoogle Scholar
  16. 16.
    Berggren K, Johansson H-O, Tjerneld F (1995) Effect of salts and the surface hydrophobicity of proteins on partitioning in aqueous two-phase systems containing thermoseparating ethylene oxide-propylene oxide copolymers. J Chrom A 718:67–79CrossRefGoogle Scholar
  17. 17.
    Dembczyński R, Białas W, Regulski K, Jankowski T (2010) Lysozyme extraction from hen egg white in an aqueous two-phase system composed of ethylene oxide–propylene oxide thermoseparating copolymer and potassium phosphate. Process Biochem 45:369–374CrossRefGoogle Scholar
  18. 18.
    Persson J, Kaul A, Tjerneld F (2000) Polymer recycling in aqueous two-phase extractions using thermoseparating ethylene oxide–propylene oxide copolymers. J Chromatogr B 743:115–126CrossRefGoogle Scholar
  19. 19.
    Abbasiliasi S, Tan JS, Ibrahim TA, Kadkhodaei S, Ng HS, Vakhshiteh F, Ajdari Z, Mustafa S, Ling TC, Rahim RA, Ariff AB (2014) Primary recovery of a bacteriocin-like inhibitory substance derived from Pediococcus acidilactici Kp10 by an aqueous two-phase system. Food Chem 151:93–100CrossRefGoogle Scholar
  20. 20.
    Kaul A, The Phase Diagram (2000) In: Hitti-Kaul R (ed) Aqueous two-phase systems: methods and protocols. Totowa, Humana Press, CrossRefGoogle Scholar
  21. 21.
    Akaraonye E, Moreno C, Knowles JC, Keshavarz T, Roy I (2012) Poly(3-hydroxybutyrate) production by Bacillus Cereus SPV using sugarcane molasses as the main carbon source. Biotechnol J 7:293–303CrossRefGoogle Scholar
  22. 22.
    Merchuk JC, Andrews BA, Asenjo JA (1998) Aqueous two-phase systems for protein separation: studies on phase inversion. J Chromatogr B 711:285–293CrossRefGoogle Scholar
  23. 23.
    Silverio SC, Rodriguez O, Tavares APM, Teixeira JA, Macedo EA (2013) Laccase recovery with aqueous two-phase systems: enzyme partitioning and stability. J Mol Catal B Enzym 87:37–43CrossRefGoogle Scholar
  24. 24.
    Pereira M, Wu YT, Venancio A, Teixeira J (2003) Aqueous two-phase extraction using thermoseparating polymer: a new system for the separation of endo-polygalacturonase. Biochem Eng J 15:131–138CrossRefGoogle Scholar
  25. 25.
    Jiang ZG, Zhang HD, Wang WT (2015) Purification of papain by metal affinity partitioning in aqueous two-phase polyethylene glycol/sodium sulfate systems. J Sep Sci 38:1426–1432CrossRefGoogle Scholar
  26. 26.
    Pandey SK, Banik RM (2011) Extractive fermentation for enhanced production of alkaline phosphatase from bacillus licheniformis MTCC 1483 using aqueous two-phase systems. Bioresour Technol 102:4226–4231CrossRefGoogle Scholar
  27. 27.
    Li LJ, Jin YR, Wang XZ, Liu Y, Wu Q, Shi XL, Li XW (2015) Ionic liquid and aqueous two-phase extraction based on salting-out coupled with high-performance liquid chromatography for the determination of seven rare ginsenosides in Xue-Sai-Tong injection. J Sep Sci 38:3055–3062CrossRefGoogle Scholar
  28. 28.
    Tou BSY, Neo KE, Tey BT, Ng MYT (2014) Effect of phase inversion and separation on hepatitis B core antigen extraction from unclarified bacterial feedstock using aqueous two-phase system. Sep Purif Technol 130:45–55CrossRefGoogle Scholar
  29. 29.
    Miao S, Chen J, Cao X (2010) Preparation of a novel thermo-sensitive copolymer forming recyclable aqueous two-phase systems and its application in bioconversion of penicillin G. Sep Purif Technol 75:156–164CrossRefGoogle Scholar
  30. 30.
    Chen B, Han J, Wang Y, Sheng C, Liu Y, Zhang G, Yan Y (2014) Separation, enrichment and determination of ciprofloxacin using thermoseparating polymer aqueous two-phase system combined with high performance liquid chromatography in milk, egg, and shrimp samples. Food Chem 148:105–111CrossRefGoogle Scholar
  31. 31.
    Cha SH, Son JH, Jamal Y, Zafar M, Park HS (2016) Characterization of polyhydroxyalkanoates extracted from wastewater sludge under different environmental conditions. Biochem Eng J 112:1–12CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  • Yoong Kit Leong
    • 1
  • Pau Loke Show
    • 1
  • John Chi-Wei Lan
    • 2
  • Hwei-San Loh
    • 3
  • Yee Jiun Yap
    • 4
  • Tau Chaun Ling
    • 5
  1. 1.Bioseparation Research Group, Department of Chemical and Environmental Engineering, Faculty of EngineeringUniversity of Nottingham Malaysia CampusSemenyihMalaysia
  2. 2.Biorefinery & Bioprocess Engineering Laboratory, Department of Chemical Engineering and Material ScienceYuan Ze UniversityTaoyuanTaiwan
  3. 3.School of Biosciences, Faculty of ScienceUniversity of Nottingham Malaysia CampusSemenyihMalaysia
  4. 4.Department of Applied Mathematics, Faculty of EngineeringUniversity of Nottingham Malaysia CampusSemenyihMalaysia
  5. 5.Institute of Biological Sciences, Faculty of ScienceUniversity of MalayaKuala LumpurMalaysia

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