Isolation and Molecular Identification of Biosurfactant Producing Soil Bacteria


  • N A N Zamani
  • T E Tengku Zainal Mulok
  • R Mat Nor
  • . .





Biosurfactant, FTIR, screening methods, surfactin, 16S rDNA gene sequencing


Biosurfactants are amphiphilic compound, having hydrophilic and hydrophobic moieties enabling them to reduce surface and interfacial tension at the surface. Their unique properties are applied in various industries such as foaming and wetting agents, emulsifiers, detergents and bioremediation. A total of 98 isolates showed biosurfactant activity using hemolytic activity, drop collapse test and oil spreading assay. All isolates were rod-shaped, Gram positive and majority of them were non-endospore former. Only the isolates showing the highest percentage of emulsification index (E24) and ability to reduce tension were used for species identification using 16S rDNA gene sequencing which were isolates A1(6) and A2(1). Both isolates were identified as Bacillus sp. cp-h50 and Bacillus sp. XT-24 respectively, rod-shaped, endospore former and Gram positive. The biosurfactant produced by both species showed high emulsification index (E24) (A1(6), 63.3% and A2(1), 46.7%) and good surfactant capacity. The size of amplified gene of 16S rDNA gene was approximately 1.5 kb. These features provide evidence that both species could be a potential biosurfactant producer with proper optimization for the production of biosurfactant. The biosurfactant produced by both bacterial species were identified as surfactin using Fourier Transform Infrared Spectroscopy (FTIR).




[1] Aziz, R. N. M., Balasaheb, S. M., Dixit, P. P. and Deshmukh, A. M. (2014). Screening of biosurfactant producing microorganisms from oil contaminated soils of Osmanabad region, Maharashtra, India. International Science Journal, 1(1), 35-39.

[2] Berry, J. D., Neeson, M. J., Dagastine, R. R., Chan, D. Y. C. and Tabor, R. F. (2015). Measurement of surface and interfacial tension using pendant drop tensiometry. Journal of Colloid and Interface Science, 454, 226-237.

[3] Dadrasnia, A. and Ismail, S. (2015). Biosurfactant production by Bacillus salmalaya for lubrication oil solubilization and biodegradation. International Journal of Environmental Research and Public Health, 12(8), 9848-9863.

[4] de Faria, A. F., Martinez, D. S. T., Barbosa, G. N. O., Vaz, B. G., Silva, I. S., Garcia, J. S., Totola, M. R., Eberlin, M. N., Grossman, M., Alves, O. L. and Durrant, L. R. (2011). Production and structural characterization of surfactin (C14/Leu7) produced by Bacillus subtilis isolate LSFM-05 grown on raw glycerol from the biodiesel industry. Process Biochemistry, 46(10), 1951-1957.

[5] Gudina, E. J., Pereira, J. F. B., Rodrigues, L. R., Coutinho, J. A. P. and Teixeira, J. A. (2012). Isolation and study of microorganisms from oil samples for application in microbial enhanced oil recovery. International Biodeterioration & Biodegradation, 68, 56-64.

[6] Hassanshahian, M. (2014). Isolation and characterization of biosurfactant producing bacteria from Persian Gulf (Bushehr provenance). Marine Pollution Bulletin, 86(1-2), 361-366.

[7] Ibrahim, M. L., Ijah, U. J. J., Manga, S. B., Bilbis,L. S. and Umar, S. (2013). Production and partial characterization of biosurfactant produced by crude oil degrading bacteria. International Biodeterioration & Biodegradation, 81, 28-34.

[8] Kannan, S. K. and Lee, K. J. (2008). Metal tolerance and antibiotic resistance of Bacillus species isolated from Sunchon Bay sediments, South Korea. Biotechnology, 7, 149-152.

[9] Korayem, A. S., Abdelhafez, A. A., Zaki, M. M. and Saleh, E. A. (2015). Optimization of biosurfactant production by Streptomyces isolated from Egyptian arid soil using Plackett-Burman design. Annals of Agricultural Sciences, 60(2), 209-217.

[10] Lopes, E. M., Castellane, T. C. L., Moretto, C., Lemos, E. G. M. and Souza, J. A. M. (2014). Emulsification properties of bioemulsifiers produced by wild-type and mutant Bradyrhizobium elkanii strains. Journal of Bioremediation and Biodegradation.

[11] Peter, J. K. and Singh, D. P. (2014). Characterization of emulsification activity of partially purified rhamnolipids from Pseudomonas fluorescens. International Journal of Innovation and Scientific Research, 3(1), 88-100.

[12] Plaza, G., Zjawiony, I. and Banat, I. (2006). Use of different methods for detection of thermophilic biosurfactant-producing bacteria from hydrocarbon-contaminated bioremediated soils. Journal of Petroleum Science and Engineering, 50(1), 71-77.

[13] Prabakaran, M. and Sumathi, R. (2014). Biosurfactant producing Pseudomonas aeruginosa and its antimicrobial activity on selected human pathogens. International Journal of Current Research in Chemistry and Pharmaceutical Sciences, 1(2), 69-78.

[14] Rufino, R. D., da Motta, S. G. N., de Luna, J. M. and Sarubbo, L. A. (2016). Conservation of the biosurfactant produced by Pseudomonas aeruginosa for environmental applications. Chemical Engineering, 49.

[15] Rufino, R. D., Luna, J. M., Campos-Takaki, G. M., Ferreira, S. R. and Sarubbo, L. A. (2012). Application of the biosurfactant produced by Candida lipolytica in the remediation of heavy metals. Chemical Engineerings, 27.

[16] Saad, S. M. I., Policova, Z. and Neumann, A. W. (2011). Design and accuracy of pendant drop methods for surface tension measurement. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 384(1-3), 442-452.

[17] Santhini, K. (2014). Isolation and screening of biosurfactant producing organisms from hydrocarbon contaminated soils from automobile workshop. International Journal of Pharmaceutical & Biological Archive, 5(2).

[18] Saravanan, V. and Vijayakumar, S. (2012). Isolation and screening of biosurfactant producing microorganisms from oil contaminated soil. J. Acad. Indus. Res, 1(5), 264-268.

[19] Shao, C., Liu, L., Gang, H., Yang, S. and Mu, B. (2015). Structural diversity of the microbial surfactin derivatives from selective esterification approach. International Journal of Molecular Sciences, 16(1), 1855-1872.

[20] Sharma, A., Soni, J., Kaur, G. and Kaur, J. (2014). A study on biosurfactant production in Lactobacillus and Bacillus sp. International Journal of Current Microbiology and Applied Sciences, 3(11), 723-733.

[21] Sriram, M. I., Gayathiri, S., Gnanaselvi, U., Jenifer, P. S., Raj, S. M. and Gurunathan, S. (2011). Novel lipopeptide biosurfactant produced by hydrocarbon degrading and heavy metal tolerant bacterium Escherichia fergusonii KLU01 as a potential tool for bioremediation. Bioresource technology, 102(19), 9291-9295.

[22] Thavasi, R., Sharma, S. and Jayalakshmi, S. (2011). Evaluation of screening methods for the isolation of biosurfactant producing marine bacteria. J Pet Environ Biotechnol.

[23] Varadavenkatesan, T. and Murty, V. R. (2013). Production of a lipopeptidenbiosurfactant by a novel Bacillus sp. and its applicability to enhanced oil recovery. ISRN Microbiology, 621519.

[24] Walter, V., Syldatk, C. and Hausmann, R. (2010). Screening concepts for the isolation of biosurfactant producing microorganisms, Adv Exp Med Biol, 672, 1-13.

[25] Youssef, N. H., Duncan, K. E., Nagle, D. P., Savage K. N., Knapp, R. M. and Mclnerney, M. J. (2004). Comparison of methods to detect biosurfactant production by diverse microorganisms. Journal of Microbiological Methods, 56, 339-347.

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How to Cite

A N Zamani, N., E Tengku Zainal Mulok, T., Mat Nor, R., & ., . (2019). Isolation and Molecular Identification of Biosurfactant Producing Soil Bacteria. International Journal of Engineering & Technology, 7(4.14), 77–81.
Received 2019-02-15
Accepted 2019-02-15
Published 2019-12-24