Abstract
Rhamnolipid is one of the most commonly used biosurfactants with the ability to reduce the surface tension of water from 72 to 30 mN/m. An indigenous isolate Pseudomonasaeruginosa S2 possessing excellent ability to produce rhamnolipid was used as a model strain to explore fermentation technology for rhamnolipid production. Using optimal medium and operating conditions (37°C, pH 6.8, and 250 rpm agitation) obtained from batch fermentation, P. aeruginosa S2 was able to produce up to 5.31 g/l of rhamnolipid from glucose-based medium. To further improve the rhamnolipid yield, a pH-stat fed-batch culture was performed by maintaining a constant pH of 6.8 through manipulating glucose feeding. The effect of influent glucose concentration on rhamnolipid yield and productivity was investigated. Using the pH-stat culture, a maximum rhamnolipid concentration (6.06 g/l) and production rate (172.5 ml/h/l) was obtained with 6% glucose in the feed. Moreover, combining pH-stat culture with fill-and-draw operation allowed a stable repeated fed-batch operation for approximately 500 h. A marked increase in rhamnolipid production was achieved, leading to the best rhamnolipid yield of approximately 9.4 g/l during the second repeated run.
Similar content being viewed by others
References
Abdel-Fattah YR, Saeed HM, Gohar YM, El-Baz MA (2005) Improved production of Pseudomonas aeruginosa uricase by optimization of process parameters through statistical experimental designs. Process Biochem 40:1707–1714
Benincasa M, Contiero J, Manresa MA, Moraes IO (2002) Rhamnolipid production by Pseudomonas aeruginosa LBI growing on soapstock as the sole carbon source. J Food Eng 54:283–288
Chang JS, Chao YP, Law WS (1998) Repeated fed-batch operations for microbial detoxification of mercury using wild-type and recombinant mercury-resistant bacteria. J Biotechnol 64:219–230
Chen CY, Baker SC, Darton RC (2006) Batch production of biosurfactant with foam fractionation. J Chem Technol Biotechnol 81:1923–1931
Fiechter A (1992) Biosurfactants: moving towards industrial application. Trends Biotechnol 10:208–217
Guerra-Santos L, Käpeli O, Fiechter A (1984) Pseudomonas aeruginosa biosurfactant production in continuous culture with glucose as carbon source. Appl Environ Microbiol 48:302–305
Guerra-Santos L, Käpeli O, Fiechter A (1986) Dependence of Pseudomonas aeruginosa continuous culture biosurfactant production on nutritional and environmental factors. Appl Microbiol Biotechnol 24:443–448
Hauser G, Karnovsky ML (1957) Rhamnose and rhamnolipid biosynthesis by Pseudomonas aeruginosa. J Biol Chem 224:91–105
Hekmat D, Bauer R, Neff V (2007) Optimization of the microbial synthesis of dihydroxyacetone in a semi-continuous repeated-fed-batch process by in situ immobilization of Gluconobacter oxydans. Process Biochem 42:71–76
Jarvis FG, Johnson MJ (1949) A glycolipipd produced by Pseudomonas aeruginosa. J Am Chem Soc 71:4124–4126
Jeong HS, Lim DJ, Hwang SH, Ha SD, Kong JY (2004) Rhamnolipid production by Pseudomonas aeruginosa immobilized in polyvinyl alcohol beads. Biotechnol Lett 26:35–39
Kim HS, Jeon JW, Kim B, Ahn CY, Oh HM, Yoon BD (2006) Extracellular production of a glycolipid biosurfactant, mannosylerythritol lipid, by Candida sp. SY16 using fed-batch fermentation. Appl Microbiol Biotechnol 70:391–396
Lang S, Wullbrandt D (1999) Rhamnose lipids—biosynthesis, microbial production and application potential. Appl Microbiol Biotechnol 51:22–32
Lee Y, Lee SY, Yang JW (1999) Production of rhamnolipid biosurfactant by fed-batch culture of Pseudomonas aeruginosa using glucose as a sole carbon source. Biosci Biotechnol Biochem 63:946–947
Maier RM, Chavez GS (2000) Pseudomonas aerugionsa rhamnolipids: biosynthesis and potential applications. Appl Microbiol Biotechnol 54:625–633
Manreas MA, Bastida J, Mercadé ME, Robert M, de Andrés C, Espuny MJ, Guinea J (1991) Kinetic studies on surfactant production by Pseudomonas aeruginosa 44T1. J Ind Microbiol 8:133–136
Mata-Sandoval J, Karns J and Torrents A (1999) High-performance liquid chromatography method for the characterization of rhamnolipids mixture produced by Pseudomonas aeruginosa UG2 on corn oil. J Chromatogr 864:211–220
Matsufuji M, Nakata K, Yoshimoto A (1997) High production of rhamnolipids by Pseudomonas aeruginosa growing on ethanol. Biotechnol Lett 19:1213–1215
Mukherjee S, Das P, Sen R (2006) Towards commercial production of microbial surfactants. Trends Biotechnol 24:509–515
Perfumo A, Banat IM, Canganella F, Marchant R (2006) Rhamnolipid production by a novel thermophilic hydrocarbon-degrading Pseudomonas aeruginosa AP02-1. Appl Microbiol Biotechnol 72:132–138
Rashedi H, Jamshidi E, Mazaheri Assadi M, Bonakdarpour B (2006) Biosurfactant production with glucose as a carbon source. Chem Biochem Eng Q 20:99–106
Reiling HE, Thanei-Wyss U, Guerra-Santos LH, Hirt R, Käppeli O, Fiechter A (1986) Pilot plant production of rhamnolipid biosurfactant by Pseudomonas aeruginosa. Appl Environ Microbiol 51:985–989
Santa Anna LM, Sebastian GV, Menezes EP, Alves TLM, Santos AS, Pereira Jr N, Freire DMG (2002) Production of biosurfactants from Pseudomonasaeruginosa PA1 isolated in oil environments. Braz J Chem Eng 19:159–166
Soberón-Chávez G, Lépine F, Déziel E (2005) Production of rhamnolipids by Pseudomonas aeruginosa. Appl Microbiol Biotechnol 68:718–725
Tashiro Y, Takeda K, Kobayashi, Sonomoto K, Ishizaki A, Yoshino S (2004) High butanol production by Clostridium saccharoperbutylacetonicum N1-4 in fed-batch culture with pH-stat continuous butyric acid and glucose feeding method. J Biosci Bioeng 4:263–268
Van GS, Sung S (2001) Biohydrogen production as a function of pH and substrate concentration. Environ Sci Technol 35:4726–4730
Van Hamme JD, Singh A, Ward OP (2006) Physiological aspects. Part 1 in a series of papers devoted to surfactants in microbiology and biotechnology. Biotechnol Adv 24:604–620
Venkata Ramana K, Karanth N (1989) Factors affecting biosurfactant production using Pseudomonas aeruginosa CFTR-6 under submerged conditions. J Chem Technol Biotechnol 45:249–257
Wei YH, Chou CL, Chang JS (2005) Rhamnolipid production by indigenous Pseudomonas aeruginosa J4 originating from petrochemical wastewater. Biochem Eng J 27:146–154
White D (1995) The physiology and biochemistry of prokaryotes, chapter 2. Oxford University Press, New York
Yeh MS, Wei YH, Chang JS (2005) Enhanced production of surfactin from Bacillussubtilis by addition of solid carriers. Biotechnol Prog 21:1329–1334
Yeh MS, Wei, Y-H, Chang JS (2006) Bioreactor design for enhanced carrier-assisted surfactin production with Bacillus subtilis. Process Biochem 41:1799–1805
Zhao YN, Chen G, Yao SJ (2006) Microbial production of 1,3-propanediol from glycerol by encapsulated Klebsiella pneumoniae. Biochem Eng J 32:93–99
Acknowledgements
The authors gratefully acknowledge the financial support from Taiwan’s Ministry of Economic Affairs under the grant number 95-EC-17-A-10-S1-013.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chen, SY., Wei, YH. & Chang, JS. Repeated pH-stat fed-batch fermentation for rhamnolipid production with indigenous Pseudomonas aeruginosa S2. Appl Microbiol Biotechnol 76, 67–74 (2007). https://doi.org/10.1007/s00253-007-0980-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-007-0980-2