• Logo
  • SBMUJournals

Production and Characterization of Biosurfactants Using Bacteria Isolated from Acidic Hot Springs

Somayeh Ebrahimi, Hasan Jalili, Khosro Khajeh, Kambiz Akbari Noghabi, Maryam Tajabadi ebrahimi, Abdeltif Amrane




Background and objective: Biosurfactants are increasingly used by food industries due to their low toxicities and unique structures. In this study, biosurfactants were produced and characterized for the first time using acidic bacteria isolated from acidic hot springs in Bushehr Province, Iran.

Material and methods: Screening and identification of the most efficient species for biosurfactant production were carried out on 12 bacterial species using several experiments such as hemolysis, surface tension, emulsification index and diameter of clear zone. In addition to biosurfactant production, kinetics, stability and structural and thermal analysis were carried out for the bacterial strains using thin layer chromatography, Fourier Transform Infrared, nuclear magnetic resonance and differential scanning calorimetry.

Results and conclusion: The biosurfactant from the selected bacteria (0.1 g l-1) was thermally stable at 120°C for 30 min. Stability at temperatures up to 140°C was confirmed using differential scanning calorimetry. The most significant novelty included the fact that the surface property was preserved until an osmolarity of 4% w v-1. Decreased surface tension and the emulsification potential were only reported at concentrations higher the highlighted concentration. Biological assay showed that Staphylococcus aureus was susceptible to produced biosurfactants, while no susceptibility was seen in Escherichia coli. Degeneration of SW480 cell line exposed to 0.601 µg µl-1 of the biosurfactant was detected after 24 h. The structural analysis showed that the biosurfactant was similar to surfactin as a food bioemulsifier.

Conflict of interest: The authors declare that they have no conflict of interest.


▪ Acidic hot spring ▪ Biosurfactant production ▪ Characterization ▪ Extremophile


Banat IM, Makkar RS, Cameotra SS. Potential commercial applications of microbial surfactants. Appl Microbiol Biotechnol. 2000;53(5):495-508.

Mukherjee AK, Das K. Microbial Surfactants and Their Potential Applications: An Overview. In: Sen R, editor. Biosurfactants. New York, NY: Springer New York; 2010. p. 54-64. 10.1007/978-1-4419-5979-9_4.

Rahman PKSM, Gakpe E. Production, characterisation and applications of biosurfactants - Review. Biotechnol. 2008;7(2):360-70. doi: 10.3923/biotech.2008.360.370.

Kazemi GA, Tajabadi Ebrahimi M, Jalili H, Dameshqyan M. Isolation, identification and considering the antagonistic properties of bacteria from hot water acidic spring in Boushehr province. New Cellular and Molecular Biotechnology Journal. 2014;4(14):65-72.

Darvishi P, Ayatollahi S, Mowla D, Niazi A. Biosurfactant production under extreme environmental conditions by an efficient microbial consortium, ERCPPI-2. Colloids Surf B Biointerfaces. 2011;84(2):292-300.

Płaza GA, Zjawiony I, Banat IM. Use of different methods for detection of thermophilic biosurfactant-producing bacteria from hydrocarbon-contaminated and bioremediated soils. J Petro Sci Eng. 2006;50(1):71-7. doi: 10.1016/j.petrol.2005.10.005.

Sen R. Response surface optimization of the critical media components for the production of surfactin. J Chem Technol Biotechnol. 1997;68(3):263-70.

Lotfabad TB, Shourian M, Roostaazad R, Najafabadi AR, Adelzadeh MR, Noghabi KA. An efficient biosurfactant-producing bacterium Pseudomonas aeruginosa MR01, isolated from oil excavation areas in south of Iran. Colloids Surf B Biointerfaces. 2009;69(2):183-93.

Jain RM, Mody K, Mishra A, Jha B. Isolation and structural characterization of biosurfactant produced by an alkaliphilic bacterium Cronobacter sakazakii isolated from oil contaminated wastewater. Carbohydrate Polymers. 2012;87(3):2320-6.

Cooper DG, Goldenberg BG. Surface-active agents from two Bacillus species. Appl Environ Microbiol. 1987;53(2):224-9.

Morikawa M, Daido H, Takao T, Murata S, Shimonishi Y, Imanaka T. A new lipopeptide biosurfactant produced by Arthrobacter sp. strain MIS38. J Bacteriol. 1993;175(20):6459-66.

Siegmund I, Wagner F. New method for detecting rhamnolipids excreted byPseudomonas species during growth on mineral agar. Biotechnol Tech. 1991;5(4):265-8.

Rosenberg M, Gutnick D, Rosenberg E. Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiol Lett. 1980;9(1):29-33.

Abbasi H, Hamedi MM, Lotfabad TB, Zahiri HS, Sharafi H, Masoomi F, et al. Biosurfactant-producing bacterium, Pseudomonas aeruginosa MA01 isolated from spoiled apples: physicochemical and structural characteristics of isolated biosurfactant. J Biosci Bioeng. 2012;113(2):211-9.

Yeh MS, Wei YH, Chang JS. Enhanced Production of Surfactin from Bacillussubtilis by addition of solid carriers. Biotechnol Prog. 2005;21(4):1329-34.

Abdel-Mawgoud AM, Aboulwafa MM, Hassouna NA-H. Characterization of surfactin produced by Bacillus subtilis isolate BS5. Appl Biochem Biotechnol. 2008;150(3):289-303.

El-Sersy NA. Plackett-burman design to optimize biosurfactant production by marine Bacillus subtilis N10. Romanian Biotechnological Letter. 2012;17(2):7049-64.

Makkar R, Cameotra SS. Effects of various nutritional supplements on biosurfactant production by a strain of Bacillus subtilis at 45 C. Journal of surfactants and detergents. 2002;5(1):11-7.

Jokari S, Rashedi H, Amoabediny G, Naghizadeh Dilmaghani S, Mazaheri Assadi M. Optimization of Surfactin Production by Bacillus subtilis ATCC 6633 in a Miniaturized Bioreactor. International Journal of Environmental Research. 2013;7(4):851-8.

Makkar R, Cameotra SS. Biosurfactant production by a thermophilic Bacillus subtilis strain. Journal of Industrial Microbiology and Biotechnology. 1997;18(1):37-42.

Lotfabad TB, Abassi H, Ahmadkhaniha R, Roostaazad R, Masoomi F, Zahiri HS, et al. Structural characterization of a rhamnolipid-type biosurfactant produced by Pseudomonas aeruginosa MR01: enhancement of di-rhamnolipid proportion using gamma irradiation. Colloids Surf B Biointerfaces. 2010;81(2):397-405.

Priya T, Usharani G. Comparative study for biosurfactant production by using Bacillus subtilis and Pseudomonas aeruginosa. Botany Research International. 2009;2(4):284-7.

Fernandes PAV, Arruda IRd, Santos AFABd, Araújo AAd, Maior AMS, Ximenes EA. Antimicrobial activity of surfactants produced by Bacillus subtilis R14 against multidrug-resistant bacteria. Braz J Microbiol. 2007;38(4):704-9.

Bauer A, Kirby W, Sherris JC, turck, Turck M. Antibiotic susceptibility testing by a standardized single disk method. American journal of clinical pathology. 1966;45(4):493.

Nozhat Z, Asadi A, Zahri S. Properties of surfactin C-15 nanopeptide and its cytotoxic effect on human cervix cancer (HeLa) cell line. Journal of Nanomaterials. 2012;2012:16.

Park SY, Kim Y. Surfactin inhibits immunostimulatory function of macrophages through blocking NK-κB, MAPK and Akt pathway. Int Immunopharmacol. 2009;9(7):886-93.

Youssef NH, Duncan KE, Nagle DP, Savage KN, Knapp RM, McInerney MJ. Comparison of methods to detect biosurfactant production by diverse microorganisms. J Microbiol Methods. 2004;56(3):339-47.

Carrillo PG, Mardaraz C, Pitta-Alvarez SI, Giulietti AM. Isolation and selection of biosurfactant-producing bacteria. World J Microbiol Biotechnol. 1996;12(1):82-4. doi: 10.1007/bf00327807.

Willumsen PA, Karlson U. Screening of bacteria, isolated from PAH-contaminated soils, for production of biosurfactants and bioemulsifiers. Biodegradation. 1996;7(5):415-23.

Suwansukho P, Rukachisirikul V, Kawai F, H-Kittikun A. Production and applications of biosurfactant from Bacillus subtilis MUV4. Sonklanakarin Journal of Science and Technology. 2008;30(1):87.

Makkar R, Cameotra SS. Production of biosurfactant at mesophilic and thermophilic conditions by a strain of Bacillus subtilis. Journal of Industrial Microbiology and Biotechnology. 1998;20(1):48-52.

Abdel-Mawgoud AM, Aboulwafa MM, Hassouna NA-H. Optimization of surfactin production by Bacillus subtilis isolate BS5. Appl Biochem Biotechnol. 2008;150(3):305-25.

Ghribi D, Ellouze-Chaabouni S. Enhancement of Bacillus subtilis lipopeptide biosurfactants production through optimization of medium composition and adequate control of aeration. Biotechnology research international. 2011;2011.

DOI: https://doi.org/10.22037/afb.v6i2.23663


  • There are currently no refbacks.