Abstract
Medium optimization for production of poly-β-hydroxyalkanoate (PHA) from Rhodobacter sphaeroides U7 cultivated in glutamate–acetate (GA) medium supplemented with 40 mM valeric acid as co-substrate under aerobic-dark condition was investigated. Studies on effect of nitrogen source and cultivation temperature by conventional and statistical methods illustrated that (NH4)2SO4 (0.2 g/l) had no effect and the optimal temperature was at 30°C. The optimum environmental conditions were found to be anaerobic-light (3000 lux) cultivation with aeration rate of 1.0 vvm and agitation speed of 200 rpm for PHA production (2.5 g/l) with the highest PHA content (65.15%) at 0.5 vvm, and 200 rpm. Under this optimized medium and condition, PHA production from R. sphaeroides U7 increased 3.86-folds (from 0.69 to 2.66 g/l) (PHA content increased 1.5-folds). The biopolymer was purified and characterized by using 13C NMR, FTIR, DSC, X-ray diffraction and intrinsic viscosity techniques to be a copolymer poly(β-hydroxybutyrate-co-β-hydroxyvalerate) (PHBV) consisting of 84.8 mol% β-hydroxybutyric acid (HB) and 15.2 mol% β-hydroxyvaleric acid (HV).
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References
Box GEP, Hunter WG, Hunter JS (1978) Statistics for experimenter. Wiley, New York
Brandl H, Gross RA, Lenz RW et al (1991) The accumulation of poly (3-hydroxyalkanoates) in Rhodobacter sphaeroides. Arch Microbiol 155:337–340
Choi JI, Lee SY (1999) High-level production of poly(3-Hydroxybutyrate-co-3-hydroxyvalerate) by fed-batch culture of recombinant Escherichia coli. Appl Environ Microbiol 65(10):4363–4368
Chua ASM, Takabatake H, Satoh H et al (2003) Production of polyhydroxyalkanoates (PHA) by activated sludge treating municipal wastewater: effect of pH, sludge retention time (SRT), and acetate concentration in influent. Water Res 37:3602–3611
Doi Y (1990) Microbial polyester. VCH Publishers, Inc., New York
Ganzeveld KJ, Hagen AV, Agteren MHV et al (1999) Upgrading of organic waste: production of the copolymer poly-3-hydroxybutyrate-co-valerate by Ralstonia eutrophus with organic waste as sole carbon source. J Clean Prod 7:413–419
Groth E, Moo-Young M, Chisti Y (1999) Fermentation optimization for the production of poly(β-hydroxybutyric acid) microbial thermoplastic. Enzyme Microb Technol 25:132–141
Grothe E, Chisti Y (2000) Poly(β-hydroxybutyric acid) thermoplastic production by Alcaligenes latus: behavior of fed-batch cultures. Bioprocess Eng 22:441–449
Hahn SK, Chang YK, Lee SY(1995) Recovery and characterization of poly(3-hydroxybutyric acid) synthesized in Alcaligenes eutrophus and recombinant Escherichia coli. Appl Environ Microbiol 61(1):34–39
Hocking PJ, Marchessault RH (1994) Chemical and technology of biodegradable polymers. Chapman and Hill, London
Ivanovsky RN, Krasilnikova EN, Berge IA (1997) A proposed citramalate cycle for acetate assimilation in the purple non-sulfur bacterium Rhodospirillum rubrum. FEMS Microbiol Lett 153(2):399–404
Jung YM, Park JS, Lee YH (2000) Metabolic engineering of Alcaligenes eutrophus through the transformation of cloned phbCAB genes for the investigation of the regulatory mechanism of polyhydroxyalkanoate biosynthesis. Enzyme Microb Technol 26:201–208
Kemavongse K, Prasertsan P, Upichit A et al (2007) Effect of co-substrate on production of poly-b-hydroxybutyrate (PHB) and copolymer PHBV from newly identified mutant Rhodobacter sphaeroides U7 cultivated under aerobic-dark condition. Songklanakarin J Sci Technol 29(4):1101–1113
Libergesell M, Hustede E, Timm A et al (1991) Formation of poly(3-hydroxyalkanoic) acids by phototrophic and lithotrophic bacteria. Arch Microbiol 155:415–421
Lorrungruang C, Martthoong J, Sasake K et al (2006) Selection of photosynthetic bacterium Rhodobacter sphaeroides 14F for polyhydroxyalkanoate production with two stage aerobic cultivation. J Biosci Bioeng 120(2):128–131
Luli GW, Strohl WR (1990) Comparison of growth, acetate production, and acetate inhibition of Escherichia coli strains in batch and fed-batch fermentations. Appl Environ Microbiol 56(4):1004–1011
Luo S, Netravali AN (2003) A study of physical and mechanical properties of poly(hydroxybutyrate-co-hydroxyvalerate) during composting. Polym Degrad Stab 80:59–66
Madmarn W, Prasertsan P, Choorit W et al (2002) Strain improvement of halotolerant photosynthetic bacteria for over production of 5-aminolevulinic acid under aerobic-darkcondition. Extended abstracts, the 3rd national symposium on graduate research 18–19 July, 2002. Suranaree University of Technology, Nakornratchasrima
Mahishi LH, Tripathi G, Rawal SK (2003) Poly(3-hydroxybutyrate) (PHB) synthesis by recombinant Escherichia coli harbouring Streptomyces aureofaciens PHB biosynthesis genes: effect of various carbon and nitrogen sources. Microbiol Res 158:19–27
Pich A, Schiemenz N, Corten C et al (2006) Preparation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) particles in O/W emulsion. Polymer 47:1912–1920
Prasertsan P, Choorit W, Suwanno S (1993) Optimization for growth of Rhodocyclus gelatinosus in seafood processing effluents. World J Microbiol Biotechnol 9:614–617
Quagliano JC, Miyazaki SS (1999) Biosynthesis of poly-beta-hydroxybutyrate and exopolysaccharides on Azotobacter chroococcum strain 6B utilizing simple and complex carbon sources. App Biochem Biotech 82(3):199–208
Quillaguaman J, Delgado O, Mattiasson B et al (2006) Poly(β-hydroxybutyrate) production by a moderate halophile, Halomonas boliviensis LC1. Enzyme Microb Technol 38:148–154
Quillaguaman J, Muñoz M, Mattiasson B et al (2007) Optimizing conditions for poly(β-hydroxybutyrate) production by Halomonas boliviensis LC1 in batch culture with sucrose as carbon source. Appl Microbiol Biotechnol 74:981–986
Reddy CSK, Ghai R, Kalia VC (2007) Polyhydroxyalkanoates : an overview. Biores Technol 87(2):137–146
Sangkharak K, Prasertsan P (2007) Optimization of polyhydroxybutyrate production from a wild type and two mutant strains of Rhodobacter sphaeroides using statistical method. J Biotechnol 132:331–340
Staley JT, Bryant MP, Pfenning N et al (1989) Bergey’s manual of systematic bacteriology, 3rd edn. The Williams and Wilkins Co., Baltimore
Sunitha K, Lee JK, Tae-Kwang O (1999) Optimization of medium components for phytase production by E. coli using response surface methodology. Bioprocess Eng 21:477–481
Tangprasittipap A, Prasertsan P (2007) Biosynthesis of intracellular 5-aminolevulinic acid by a newly identified halotolerant R. sphaeroides. Biotech Lett 29(5):773–778
Yigit DO, Gunduz U, Turker L et al (1999) Identification of by-products in hydrogen producing bacteria; Rhodobacter sphaeroides O.U. 001 grown in the waste water of a sugar refinery. J Biotechnol 70:125–131
Yuksekdag ZN, Aslim B, Beyatli Y et al (2004) Effect of carbon and nitrogen sources and incubation times on poly-beta-hydroxybutyrate (PHB) synthesis by Bacillus subtilis 25 and Bacillus megaterium 12. Afr J Biotechnol 3(1):63–66
Acknowledgment
This research work was financially supported by Master Research Grants (MAG) of Thailand Research Fund (TRF), Department of Commission on Higher Education, Graduate School and Faculty of Agro-Industry of Prince of Songkla University.
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Kemavongse, K., Prasertsan, P., Upaichit, A. et al. Poly-β-hydroxyalkanoate production by halotolerant Rhodobacter sphaeroides U7. World J Microbiol Biotechnol 24, 2073–2085 (2008). https://doi.org/10.1007/s11274-008-9712-8
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DOI: https://doi.org/10.1007/s11274-008-9712-8