Abstract
A new halogen-free and environmental-friendly method using water and ethanol is developed as an alternative for the recovery of polyhydroxyalkanoates (PHA) from recombinant Cupriavidus necator in comparison to the established chloroform extraction method. After optimisation, our results showed that the halogen-free method produced a PHA with 81% purity and 96% recovery yield, in comparison to the chloroform extraction system which resulted in a highly pure PHA with 95% yield. Although the purity of the PHA using the new method is lower, the molecular weight of the extracted PHA is not compromised. This new method can be further developed as an alternative and more environmental-friendly method for industrial application.
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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, Malaysia. Bioresource Technol 99:4903–4909
Chen Y, Xu Q, Yang H, Gu G (2001) Effects of cell fermentation time and biomass drying strategies on the recovery of poly-3-hydroxyalkanoates from Alcaligenes eutrophus using a surfactant-chelate aqueous system. Proc Biochem 36:773–779
Choi J, Lee SY (1999) Efficient and economical recovery of poly(3-hydroxybutyrate) from recombinant Escherichia coli by simple digestion with chemicals. Biotechnol Bioeng 62:546–553
de Koning GJM, Witholt B (1997) A process for the recovery of poly (hydroxyalkanoates) from Pseudomonads Part 1: solubilization. Bioproc Biosyst Eng 17:7–13
Ghatnekar MS, Pai JS, Ganesh M (2002) Production and recovery of poly-3-hydroxybutyrate from Methylobacterium sp V49. J Chem Tech Biotech 77:444–448
Hassan MA, Mohammadi M, Abd Razak NA, Chong ML, Shirai Y, Yunus WMZ, Suraini AA, Amirul AAA, Hasfalina CM, Syairah A, Phang LY (2010) A method of recovering an intracellular polyhydroxyalkanoate (PHA). PCT/MY2010/000333
Hejazi P, Vasheghani-Farahani E, Yamini Y (2003) Supercritical fluid disruption of Ralstonia eutropha for poly(β-hydroxybutyrate) recovery. Biotechnol Progr 19:1519–1523
Jiang X, Ramsay JA, Ramsay BA (2006) Acetone extraction of mcl-PHA from Pseudomonas putida KT2440. J Microbiol Methods 67:212–219
Kapritchkoff FM, Viotti AP, Alli RCP, Zuccolo M, Pradella JGC, Maiorano AE, Miranda EA, Bonomi A (2006) Enzymatic recovery and purification of polyhydroxybutyrate produced by Ralstonia eutropha. J Biotechnol 122:453–462
Kurdikar DL, Strauser FE, Solodar AJ, Paster MD, Asrar J (2000a) Methods of PHA extraction and recovery using non-halogenated solvents. US006043063A
Kurdikar DL, Strauser FE, Solodar AJ, Paster MD (2000b) High temperature PHA extraction using PHA-poor solvents. US6087471
Mantelatto PE, Durao NA (2008) Process for extracting and recovering polyhydroxyalkanoates (Phas) from cellular biomass. US20080193987A1
Mantelatto PE, Duzzi AM, Sato T, Durao NA, Nonato RV, Rocchiccioli C, Kesserlingh SM (2005) Process for recovering polyhydroxialkanoates (‘Phas’) from cellular biomass. WO/2005/052175A2
Mattingly SJ, Best GK (1971) The effect of temperature on lysis of cells and cell walls of Bacillus psychrophilus. Can J Microbiol 17:1161–1168
Mohammadi M, Hassan MA, Shirai Y, Hasfalina CM, Ariffin H, Lian- Ngit Y, Mumtaz T, Chong ML, Phang LY (2011a) Separation and purification of polyhydroxyalkanoates from newly isolated Comamonas sp. EB172 by simple digestion with sodium hydroxide. Separ Sci Technol (in press)
Mohammadi M, Hassan MA, Phang LY, Shirai Y, Hasfalina CM, Ariffin H, Amirul AAA, Syairah A (2011b) Efficient PHA recovery from recombinant Cupriavidus necator by using low concentration of NaOH. Environ Eng Sci (in press)
Mumtaz T, Suraini AA, Abdul Rahman NA, Phang LY, Shirai Y, Hassan MA (2009) Fed-batch production of P(3HB-co-3HV) copolymer by Comamonas sp EB 172 using mixed organic acids under dual nutrient limitation. Eur J Sci Res 33:374–384
Nair AJ (2008) Principles of biotechnology. Laxmi Publications, New Delhi
Page WJ, Cornish A (1993) Growth of Azotobacter vinelandii UWD in fish peptone medium and simplified extraction of poly-β-hydroxybutyrate. Appl Environ Microbiol 59:4236–4244
Peters KE, Walters CC, Moldowan JM (2005) The biomarker guide: biomarkers and isotopes in the environment and human history. Cambridge University Press, Cambridge
Tortora GJ (2010) Microbiology an introduction, 10th edn. Pearson Benjamin Cummings, San Francisco
Yang YH, Brigham C, Willis L, Rha C, Sinskey A (2011) Improved detergent-based recovery of polyhydroxyalkanoates (PHAs). Biotechnol Lett 33:937–942
Acknowledgments
The financial and technical support received from the Federal Land Development Authority (FELDA), Ministry of Science Technology and Innovation (MOSTI), Malaysia and Japan Society for Promotion of Science (JSPS), Japan are gratefully acknowledged. The authors are extremely grateful to Dr. Sudesh of Universiti Sains Malaysia, Dr. Mei Ling Chong of Technology Park Malaysia, Dr. Tabassum Mumtaz and Ms. Lian Ngit Yee of Universiti Putra Malaysia for their generous support during the experiments.
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Mohammadi, M., Hassan, M.A., Phang, LY. et al. Recovery and purification of intracellular polyhydroxyalkanoates from recombinant Cupriavidus necator using water and ethanol. Biotechnol Lett 34, 253–259 (2012). https://doi.org/10.1007/s10529-011-0783-5
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DOI: https://doi.org/10.1007/s10529-011-0783-5