Production of rhamnolipid (a biosurfactant) using free and immobilized cells of Pseudomonas sp.

Bu çalışmada, bir büyosürfektan olan ramnolipidin Pseudomonas sp.’den üretimi araştırılmıştır. Nutrient agarda üretilen Pseudomonas sp. örnekleri sıvı besiyerine aktarılmıştır. 24 saatlik inkübasyon sonrası, 2 ml’lik kültür örnekleri ramnolipid üretimi için, farklı bir sıvı besiyerine aktarılarak 26. saatte sonuçlar gözlenmiştir. Çalışmada, ayrıca ramnolipid üretimine sıcaklığın, pH’ın ve glukoz konsantrasyonunun etkisi de incelenmiştir. İmmobilize (tutuklanmış) hücreler içinde aynı metotlar uygulanmıştır. Ramnolipid üretimi, serbest hücrelerde tutuklanmış hücrelerden daha yüksek bulunmuştur. Serbest hücreler, etkin ramnolipid üretimi için kullanılabilirler.

Anahtar Kelimeler:

Pseudomonas, glukoz, lipit, pH, sıcaklık

Serbest ve tutuklanmış Pseudomonas sp. hücrelerinden ramnolipid (biyosürfektan) eldesi

This study presents a method for the production of rhamnolipid, a biosurfactant, by Pseudomonas sp. Pseudomonas sp. cells that were grown in nutrient agar were inoculated into sterile liquid medium. Following an incubation period of 24 h, 2 ml of cells were inoculated into a different liquid medium and the results were obtained at the end of 26 hours incubation time. In our study, the effects of temperature, pH, and glucose concentration on rhamnolipid production were also investigated. Later, the same procedure was applied to immobilized cells that were kept away from the free microorganisms. The production of rhamnolipid by free cells was found to be much higher than that of immobilized cells. Free cells could be used for rhamnolipid production effectively.

Keywords:

Pseudomonas, glucose, lipids, pH, temperature,

PDF

___

1. Koch KA, Kappeli O, Fiechter A, Reiser J: Hydrocarbon assimilation and biosurfactant production in Pseudomonas aeruginosa mutants. J Bacteriol, 173, 4212- 4219, 1991.
2. Reiser J, Koch AK, Jeny K, Kappeli O: Structure, properties and production of biosurfactants. In, Obringer JW, Tillinghast HS (Eds): Advances in Biotechnology for Aerospace Applications. pp. 85- 97, Portfolio, The Woodlands, 1989.
3. Prieto LM, Michelon M, Burkert JFM, Kalil SJ, Burkert CAV: The production of rhamnolipid by a Pseudomonas aeruginosa strain isolated from a southern coastal zone in Brazil. Chemosphere,71, 1781-1785, 2008.
4. Singer MEV, Finnerty WR: Physiology of biosurfactant synthesis by Rhodococcus Species H13-A. Canadian J Microbiol, 36, 741-745, 1990.
5. Costa SGAO, Nitschke M, Haddad R, Eberlin, MN, Contiero J: Production of Pseudomonas aeruginosa LBI rhamnolipids following growth on Brazilian native oils. Process Biochem, 41, 483-488, 2006.
6. Yilmaz EŞ, Sidal U: Investigation of antimicrobial effects of a Pseudomonas-originated biosurfactant. Biologia, 60, 723-725, 2005.
7. Champagne CP, Gaudy C, Poncelet D, Neufeld RJ: Lactobacillus lactis Release from calcium alginate beads. Appl and Environ Microbiol, 58, 1429-1434, 1992.
8. Cheetham PSJ, Blunk KW, Bucke C: Physical studies on cell immobilization using calcium alginate gels. Biotech Bioeng, 21, 2155- 2168, 1979.
9. Guoqlang D, Kaul R, Mattiasson B: Evaluation of alginate-immobilized Lactobacillus casei for lactate production. Appl Microbiol Biotechnol, 36, 309- 314, 1991.
10. Hang YD, Hamamci H, Woodams EE: Production of L (+)- lactic acid by Rhizopus oryzae immobilized in calcium alginate gels. Biotechnology Letters, 11, 119-120, 1989.
11. Para JL, Guinea J, Manresa MA, Robert M, Mercade ME, Comelles F, Bosch MP: Chemical characterization and physicochemical behavior of biosurfactants. J Am Oil Chem Society, 66, 141-145,1989.
12. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F: Colorimetric method of determination of sugar and related substances. Anal Chem, 28, 350-356,1956.
13. Miller GL: Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem, 31, 426- 428, 1959.
14. Mercade ME, Manresa MA: The use of agroindustrial by products for biosurfactant production. J Amr Oil Chem Society, 71, 61-64, 1994.
15. Mercade ME, Manresa MA, Robert M: Olive Oil Mill Effluent (OOME) New substrate for biosurfactant production. Bioresource Technology, 43, 1- 6, 1993.
16. Ramana KV, Karanth NG: Production of biosurfactants by the resting cells of Pseudomonas aeruginosa CFTR- 6. Biotech Lett, 11, 437- 442, 1989.
17. Sidal U, Kurtonur C, Kolankaya N: Pseudomonas sp. ile zeytinyağı fabrikası atığından biyosürfektan eldesi. Tr J Biology, 24, 611-625, 2000.
18. Rocha C, San Blas F, San Blas G, Vierma L: Biosurfactant production by two isolates of Pseudomonas aeruginosa. World J Microbiol Biotechn, 8, 125-128, 1992.
19. Ramana KV, Charyulu NCLN, Karanth NG: A mathematical model for the production of biosurfactants by Pseudomonas aeruginosa CFTR-6: Production of Biomass. J Chem Tech Biotec, 51, 525-538, 1991.
20. Rosenberg E: Exploiting microbial growth on hydrocarbons-new markets. Tıb Tech October, 11, 419-424, 1993.
21. Jamuna R, Sai PST, Vora S, Ramakrishna SV: Optimization of critical parameters for immobilization of yeast cells to alginate gel matrix. J Ferm Bioeng, 73, 319-322, 1992.
22. Kanwar SS, Chadha BS, Tewari HK, Sharma VK: Continuous production of lactic acid from molasses by free and immobilized Sporolactobacillus cellulosolvens. World J Microbiol Biotech, 11, 687- 688, 1995.
23. Goksungur Y, Güvenç U: Batch and continuous production of lactic acid from beet molasses by Lactobacillus delbrueckii IFO 3202. J Chem Tech Biotechnol, 69, 399-404, 1997.
24. Abu-Ruwaida AS, Banat IM, Hadidirto S, Salem A, Kadri M: Isolation of biosurfactant-producing bacteria, product characterization and evaluation. ACTA Biotechnologica, 11, 315-324, 1991.
25. Jarvis FG, Johnson MJ: A glycolipide produced by Pseudomonas aeruginosa. J Am Chem Soc, 71, 4124- 4126, 1949.
26. Hauser G, Karnovsky ML: Studies on the production of glycolipide by Pseudomonas aeruginosa. J Bacteriol, 68, 645- 654, 1954.
27. Hauser G, Karnovsky ML: Rhamnose and rhamnolipide biosynthesis by Pseudomonas aeruginosa. J Biol Chem, 224, 91-105, 1957.
28. Hauser G, Karnovsky ML: Studies on the biosynthesis of L-rhamnose. J Biol Chem, 233, 287- 291, 1958.
29. Guerra-Santos L, Kappeli O, Fiechter A: Pseudomonas aeruginosa biosurfactant production in continuous culture with glucose as carbon source. Appl Environ Microbiol, 48, 301-305, 1984.
30. Abouseoud M, Yataghene A, Amrane A, Maachi R: Biosurfactant production by free and alginate entrapped cells of Pseudomonas fluorescens. J Ind Microbiol Biotechnol, 35, 1303-1308, 2008.