Preliminary Study of SurfactinProduction by Malaysian Local Isolates of Bacillus Subtilis


  • Saed Abdullah Hasan Department of Therapeutic nutrition, Higher institute of Science And Technology, Al -Bayda-Libya
  • Ahmed M A Hamad Department of Therapeutic nutrition, Higher institute of Science And Technology, Al -Bayda-Libya
  • Hana S. Mohammad Department of chemistry Faculty of science, Omar Al-Mukhtar university –Libya
  • Amena. A. Abdulrazeg Department of Nursing, Higher institute of Science And Technology, Cyrene- Libya



Bacillus subtilis, Surfactin , Production , High-Performance Liquid Chromatography (HPLC),, Bio-Spectrophotometer


Surfactin is one of the most powerful lipopeptidebiosurfactants produced by various strains of Bacillus subtilis. It has exceptional surface activity, with antiviral, antibacterial, and antitumor properties. The four local isolates, which were named Bacillus subtilis1M, 3M, 7M, and 8M were provided by the School of Biosciences and Biotechnology, Faculty of Science and Technology, University of Kebangsaan, Malaysia. In this study, fermentation on shaker flasks was carried out to assess the ability of four local isolates of Bacillus subtilis strains to produce surfactin by using Cooper’s media formulation, and comparing their production with a commercial strain of Bacillus subtilis ATCC 21332, which was obtained from the American Type Culture Collection. High-Performance Liquid Chromatography (HPLC) was used for surfactin identification and surfactin concentration measurements. Results obtained show the four local isolates have the ability to produce surfactin. The Bacillus subtilis3M strain showed the highest amount of surfactin production with 117 (3) mg/L, while the Bacillus subtilis1M strain produced the lowest amount with 65 (5.4) mg/L. In addition, the production of Bacillus subtilis ATCC 21332 strain was found at 101(4) mg/L under the same fermentation conditions.


Download data is not yet available.


Abdel-Mawgoud, A. M., Aboulwafa, M. M., & Hassouna, N. A.-H. (2008). Characterization of surfactin produced by Bacillus subtilis isolate BS5. Applied Biochemistry and Biotechnology, 150, 289-303.

Abushady, H., Bashandy, A., Aziz, N., & Ibrahim, H. (2005). Molecular characterization of Bacillus subtilis surfactin producing strain and the factors affecting its production. International Journal of Agriculture and Biology, 7(3), 337-344.

Al-Bahry, S., Al-Wahaibi, Y., Elshafie, A., Al-Bemani, A., Joshi, S., Al-Makhmari, H., & Al-Sulaimani, H. (2013). Biosurfactant production by Bacillus subtilis B20 using date molasses and its possible application in enhanced oil recovery. International Biodeterioration & Biodegradation, 81, 141-146.

Amit, P., & Rukhsar, A. (2013). Optimization and characterization of biosurfactant producing microbes isolated from oil contaminated soil and expression of biosurfactant genes in E. coli. International Journal of Pharmaceutical Research and Bio-science, 2(2), 46-67.

Campos, J. M., Montenegro Stamford, T. L., Sarubbo, L. A., de Luna, J. M., Rufino, R. D., & Banat, I. M. (2013). Microbial biosurfactants as additives for food industries. Biotechnology progress, 29(5), 1097-1108.

Chen, H.-L., Chen, Y.-S., & Juang, R.-S. (2008). Flux decline and membrane cleaning in cross-flow ultrafiltration of treated fermentation broths for surfactin recovery. Separation and purification technology, 62(1), 47-55.

Cheng, Y. H., Zhang, N., Han, J. C., Chang, C. W., Hsiao, F. S. H., & Yu, Y. H. (2018). Optimization of surfactin production from Bacillus subtilis in fermentation and its effects on Clostridium perfringens‐induced necrotic enteritis and growth performance in broilers. Journal of Animal Physiology and Animal Nutrition, 102(5), 1232-1244.

dos Santos, S. C., Fernandez, L. G., Rossi-Alva, J. C., & de Abreu Roque, M. R. (2010). Evaluation of substrates from renewable-resources in biosurfactants production by Pseudomonas strains. African journal of biotechnology, 9(35).

Fahle, A., Bereswill, S., & Heimesaat, M. M. (2022). Antibacterial effects of biologically active ingredients in hop provide promising options to fight infections by pathogens including multi-drug resistant bacteria. European Journal of Microbiology and Immunology, 12(1), 22-30.

Isa, M. H. M., Coraglia, D. E., Frazier, R. A., & Jauregi, P. (2007). Recovery and purification of surfactin from fermentation broth by a two-step ultrafiltration process. Journal of Membrane Science, 296(1-2), 51-57.

Jajor, P., Piłakowska-Pietras, D., Krasowska, A., & Łukaszewicz, M. (2016). Surfactin analogues produced by Bacillus subtilis strains grown on rapeseed cake. Journal of Molecular Structure, 1126, 141-146.

Jiao, S., Li, X., Yu, H., Yang, H., Li, X., & Shen, Z. (2017). In situ enhancement of surfactin biosynthesis in Bacillus subtilis using novel artificial inducible promoters. Biotechnology and Bioengineering, 114(4), 832-842.

Kinsinger, R. F., Kearns, D. B., Hale, M., & Fall, R. (2005). Genetic requirements for potassium ion-dependent colony spreading in Bacillus subtilis. Journal of bacteriology, 187(24), 8462-8469.

Li, Y., Yang, S., & Mu, B. (2010). The surfactin and lichenysin isoforms produced by Bacillus licheniformis HSN 221. Analytical letters, 43(6), 929-940.

Mulligan, C. N., Chow, T. Y.-K., & Gibbs, B. F. (1989). Enhanced biosurfactant production by a mutant Bacillus subtilis strain. Applied microbiology and biotechnology, 31, 486-489.

Pereira, J. F., Gudiña, E. J., Costa, R., Vitorino, R., Teixeira, J. A., Coutinho, J. A., & Rodrigues, L. R. (2013). Optimization and characterization of biosurfactant production by Bacillus subtilis isolates towards microbial enhanced oil recovery applications. Fuel, 111, 259-268.

Ríos, F., Fernández-Arteaga, A., Lechuga, M., & Fernández-Serrano, M. (2017). Ecotoxicological characterization of polyoxyethylene glycerol ester non-ionic surfactants and their mixtures with anionic and non-ionic surfactants. Environmental Science and Pollution Research, 24, 10121-10130.

Sousa, M., Dantas, I., Feitosa, F., Alencar, A., Soares, S., Melo, V., Gonçalves, L., & Sant'ana, H. (2014). Performance of a biosurfactant produced by Bacillus subtilis LAMI005 on the formation of oil/biosurfactant/water emulsion: study of the phase behaviour of emulsified systems. Brazilian Journal of Chemical Engineering, 31, 613-623.

Sumi, C. D., Yang, B. W., Yeo, I.-C., & Hahm, Y. T. (2015). Antimicrobial peptides of the genus Bacillus: a new era for antibiotics. Canadian journal of microbiology, 61(2), 93-103.

Vater, J., Kablitz, B. r., Wilde, C., Franke, P., Mehta, N., & Cameotra, S. S. (2002). Matrix-assisted laser desorption ionization-time of flight mass spectrometry of lipopeptide biosurfactants in whole cells and culture filtrates of Bacillus subtilis C-1 isolated from petroleum sludge. Applied and Environmental Microbiology, 68(12), 6210-6219.

Waewthongrak, W., Leelasuphakul, W., & McCollum, G. (2014). Cyclic lipopeptides from Bacillus subtilis ABS–S14 elicit defense-related gene expression in citrus fruit. PloS one, 9(10), e109386.

Wei, Y.-H., & Chu, I.-M. (2002). Mn2+ improves surfactin production by Bacillus subtilis. Biotechnology Letters, 24(6), 479-482.

Wei, Y.-H., Wang, L.-F., Changy, J.-S., & Kung, S.-S. (2003). Identification of induced acidification in iron-enriched cultures of Bacillus subtilis during biosurfactant fermentation. Journal of bioscience and bioengineering, 96(2), 174-178.

Zhi, Y., Wu, Q., & Xu, Y. (2017). Production of surfactin from waste distillers’ grains by co-culture fermentation of two Bacillus amyloliquefaciens strains. Bioresource Technology, 235, 96-103.

Zweers, J. C., Barák, I., Becher, D., Driessen, A. J., Hecker, M., Kontinen, V. P., Saller, M. J., Vavrová, L. u., & van Dijl, J. M. (2008). Towards the development of Bacillus subtilis as a cell factory for membrane proteins and protein complexes. Microbial cell factories, 7, 1-20.




How to Cite

Hasan, S. A., Hamad, A. M. A., Mohammad, H. S., & Abdulrazeg, A. A. (2023). Preliminary Study of SurfactinProduction by Malaysian Local Isolates of Bacillus Subtilis. Al-Mukhtar Journal of Sciences, 38(1), 1–8.



Research Articles