Sains Malaysiana 42(5)(2013): 615–623

 

Screening and Optimization of Biosurfactant Production by the Hydrocarbon-Degrading Bacteria

(Penyaringan dan Pengoptimuman Biosurfaktan yang Dihasilkan oleh Bakteria Pendegradasi-Hidrokarbon)

 

Ainon Hamzah*, Noramiza Sabturani & Shahidan Radiman

School of Biosciences and Biotechnology, Faculty of Science and Technology

Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor D.E. Malaysia

 

Diserahkan: 13 Julai 2011/Diterima: 24 Oktober 2012

 

ABSTRACT

Biosurfactants are amphiphilic compounds produced by microorganisms as secondary metabolite. The unique properties of biosurfactants make them possible to replace or to be added to synthetic surfactants which are mainly used in food, cosmetics and pharmaceutical industries and in environmental applications. In this study twenty hydrocarbon-degrading bacteria were screened for biosurfactant production. All of the bacterial isolates were grown in mineral salt medium (MSM) with addition of 1% (v/v) Tapis crude oil as carbon source. The presence of biosurfactant was determined by the drop-collapse test, microplate analysis, oil spreading technique, emulsification index (%EI24) and surface tension measurement. Only one isolate, Pseudomonas aeruginosa UKMP14T, was found to be positive for all the qualitative tests and reducing the surface tension of the medium to 49.5 dynes/cm with emulsification index of 25.29%. This isolate produced biosurfactant optimally at pH9.0 and incubation temperature of 37°C. Furthermore, P. aeruginosa UKMP14T when grown in MSM with addition of 1% (v/v) glycerol and 1.3 g/L ammonium sulphate with C/N ratio 14:1 produced biosurfactant with percentage of surface tension reduction at 55% or 30.6 dynes/cm with %EI24 of 43%. This percentage of surface tension reduction represents an increasing reduction in surface tension of medium by 39% over the value before optimization. This study showed that P. aeruginosa UKMP14T has the ability to biodegrade hydrocarbon and concurrently produce biosurfactant.

 

Keywords: Biosurfactant; hydrocarbon-degrading bacteria; optimization; surface tension

 

ABSTRAK

Biosurfaktan adalah sebatian ampifilik yang dihasilkan oleh mikroorganisma sebagai metabolit sekunder. Ciri-ciri biosurfaktan yang unik membolehkan mereka menggantikan atau ditambahkan kepada surfaktan sintetik yang kebanyakannya digunakan dalam industri makanan, kosmetik dan farmaseutikal dan diaplikasi dalam sekitaran. Dalam kajian ini sebanyak 20 pencilan bakteria-pencurai hidrokarbon disaring untuk penghasilan biosurfaktan. Kesemua pencilan bakteria dihidupkan di dalam medium garam mineral (MSM) yang ditambah dengan 1% (i/i) minyak mentah Tapis sebagai sumber karbon. Kehadiran biosurfaktan ditentukan dengan ujian titisan-runtuh, analisis mikroplat, teknik sebaran minyak, indeks emulsifikasi (%EI24) dan pengukuran ketegangan permukaan. Hanya satu pencilan iaituPseudomonas aeruginosa UKMP14T yang memberikan hasil positif ke atas semua ujian kualitatif dan mengurangkan ketegangan permukaan medium sehingga 49.5 dynes/cm dengan indeks emulsifikasi 25.29%. Pencilan ini menghasilkan biosurfaktan yang optimum pada pH9.0 dan suhu eraman 37°C. Selain itu, apabilaP. aeruginosa UKMP14T dihidupkan di dalam MSM dengan penambahan 1% (i/i) gliserol dan 1.3 g/L ammonium sulfat dengan nisbah C/N 14:1, ia menghasilkan biosurfaktan dengan peratus pengurangan ketegangan permukaan sebanyak 55% atau 30.6 dynes/cm dengan %EI24, 43%. Peratus pengurangan ketegangan permukaan ini mewakili peningkatan pengurangan dalam ketegangan permukaan medium sebanyak 39% jika dibandingkan sebelum pengoptimuman. Kajian ini menunjukkan bahawa P. aeruginosa UKMP14T mempunyai keupayaan untuk biodegradasi hidrokarbon serta menghasilkan biosurfaktan.

 

Kata kunci: Bakteria pencurai-hidrokarbon; biosurfaktan; ketegangan permukaan; pengoptimuman

RUJUKAN

Bouchez, M., Blanchet, D. & Vandacasteele, J.P. 1995. Degradation of polycyclic aromatic hydrocarbons by pure strains and defined strain associations: Inhibition phenomena and cometabolism. Applied Microbiology Biotechnology 43:156-164.

Bodour, A. & Miller-Maier, R.M. 1998. Application of a modified drop collapse technique for surfactant quantification and screening of biosurfactant-producing microorganisms. Journal of Microbiological Methods 32: 273-280.

Cameotra, S.S. & Makkar, R.S. 2004. Recent applications of biosurfactants as biological and immunological molecules. Current Opinion in Microbiology 7: 262-266.

Chen, C-Y., Baker, S.C. & Darton, R.C. 2007. The application of a high throughput analysis method for the screening of potential biosurfactants from natural resources. Journal of Microbiological Methods 70: 503-510.

Chen, C-Y., Lu, B-W., Wei, Y-H., Chen, W-M. & Chang, J-S. 2007. Improved production of biosurfactant with newly isolated Pseudomonas aeruginosaS2. Biotechnology Process 23(3): 661-666.

Cooper, D.G. & Goldenberg, B.G. 1987. Surface-active agents from two Bacillus species. Applied Environmental Microbiology 53: 224-229.

Hamzah, A., Rabu, A., Azmy, R.F.H.R. & Yussoff, N.A. 2010. Isolation and characterization of bacteria degrading Sumandak and South Angsi oils. Sains Malaysiana39(2): 161-168.

Kosaric, N. 1993. Biosurfactants: Production, Properties and Application. New York: Marcel Dekker, Inc.

Lotfabad, T.B., Sourian, M., Roostazad, R., Najafabadi, A.R., Adelzadeh, M.R. & Noghabi, K.A. 2009. An efficient biosurfactant-producing bacterium Pseudomonas aeruginosaMR01, isolated from oil excavation areas in south of Iran. Colloids and Surfaces B: Biointerfaces 69: 183-193.

Morikawa, M., Hirata, Y. & Imanaka, T. 2000. A study on the structure–function relationship of the lipopeptide biosurfactants. Biochimica et Biophysica Acta1488: 211-218.

Mukred, A.M., Hamid, A.A., Hamzah, A. & Yusoff, W.W.M. 2008. Enhancement of biodegradation of crude petroleum-oil in contaminated water by the addition of nitrogen sources. Pakistan Journal of Biological Sciences 11(17): 2122-2127.

Nur Faizah Abu Bakar. 2010. Biodegradation study of Tapis crude oil and condensate Terengganu by selected bacteria. MSc. Thesis, Universiti Kebangsaan Malaysia (unpublished).

Pacwa-Plociniczak, M., Plaza, G.A., Piotrowska-Seget, Z. & Cameotra, S.S. 2011. Environmental applications of biosurfactants: Recent advances. International Journal Molecular Science 12: 633-654.

Pekdemir, T., Copur, M. & Urum, K. 2005. Emulsification of crude oil–water systems using biosurfactants. Process Safety Environmental Protection 83(B1): 38-46.

Plaza, G.A., Zjawiony, I. & Banat, I.M. 2006. Use of different methods for detection of thermophilic biosurfactant-producing bacteria from hydrocarbon-contaminated and bioremediated soils. Journals of Petroleum Science and Engineering 50: 71-77.

Praveesh, B.V., Soniyambang, A.R., Mariappan, C., Kavithakumari, P., Pataniswammy, M. & Lalitha, S. 2011. Biosurfactant production by Pseudomonas sp from soil using whey as carbon source, New York Science Journal 4(4): 99-103.

Prieto, L.M., Michelon, M., Burkert, J.F.M., Kalil, S.J. & Burkert, C.A.V. 2008. The production of rhamnolipid by a Pseudomonas aeruginosastrain isolated from a southern coastal zone in Brazil. Chemosphere 71: 1781-1785.

Pornsunthorntawee, N., Arttaweeporn, N., Paisanjit, S., Somboonthanate, P., Abe, M., Rujiravanit, R. & Chavadej, S. 2008. Isolation and comparison of biosurfactants produced by Bacillus subtilis PT2 and Pseudomonas aeruginosaSP4 for microbial surfactant-enhanced oil recovery. Biochemical Engineering Journal 42: 172-179.

Silva, S.N., Farias, C.B., Rufino, R.D., Luna, J.M. & Sarubbo, L.A. 2010. Glycerol as substrate for the production of biosurfactant by Pseudomonas aeruginosaUCP0992. Colloids Surf B Biointerfaces79(1): 174-183.

Rodrigues, L., Banat, I.M., Teixeira, J. & Oliveira, R. 2006. Biosurfactants: Potential applications in medicine. Journal of Antimicrobial Chemotherapy 57: 609-618.

Wei, Y-H., Lai, C-C. & Chang, J-S. 2007. Using Taguchi experimental design methods to optimize trace element composition for enhanced surfactin production by Bacillus subtilis ATCC 21332. Process Biochemistry 42: 40-45.

Wei, Y-H., Cheng, C-L., Chien, C-C. & Wan, H-M. 2008. Enhanced di-rhamnolipid production with an indigenous isolate Pseudomonas aeruginosaJ16. Process Biochemistry 43: 769-774.

Williams, K. 2009. Biosurfactants for cosmetic application: Overcoming production challenges. MMG 445 Basic Biotechnology 5: 78-83.

Willumsen, P.A.E. & Karlson, U. 1997. Screening of bacteria, isolated from PAH-contaminated soils, for production of biosurfactants and bioemulsifiers. Biodegradation 7: 415-423.

Wu, J-Y., Yeh, K-L., Lu, W-B., Lin, C-L. & Chang, J-S. 2008. Rhamnolipid production with indigenous Pseudomonas aeruginosaEM1 isolated from oil-contaminated site. Bioresource Technology 99: 1157-1164.

Yin, H., Qjang, J., Jia, Y., Ye, J., Peng, H., Qin, H., Zhang, N. & He, B. 2009. Characteristics of biosurfactant produced by Pseudomonas aeruginosaS6 isolated from oil-containing wastewater. Process Biochemistry 44: 302-308.

Youssef, N., Duncan, K.E. & Savage, K.N. 2004. Comparison of methods to detect biosurfactant production by diverse microorganisms. Journal Microbiology Methods 56: 339-347.

Zajic, E. & Supplison, B. 1972. Emulsification and degradation of “Bunker C” fuel oil by microorganisms. Biotechnology and Bioengineering 14: 331-343.

Zhang, G-L., Wu, Y-T., Qian, X-P. & Meng, Q. 2005. Biodegradation of crude oil by Pseudomonas aeruginosa in the presence of rhamnolipid. Journal of Zhejiang University, Science B 6B(8): 725-730.

 

*Pengarang untuk surat-menyurat; email: antara@ukm.my


 

 

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