Original Article

Investigation of Physicho-chemical Properties and Characterization of Produced Biosurfactant by Selected Indigenous Oil-degrading Bacterium

Abstract

Background: Due to the amphipathic properties of biosurfactants which act on surfaces and interfaces interest by a variety of industries such as cosmetic, pharmaceutical, bioremediation and petroleum-related industries has recently increased.

Methods: Detection of a high-efficiency biosurfactant producer using preliminary screening methods from soil contaminated with crude oil was carried at the Microbiology Laboratory at Shahid Beheshti University, Tehran, Iran in 2013. Then after characterization of some physico-chemical properties of produced biosurfactant and production optimization conditions, processes of purification and complete identification were done.

Results: Pseudomonas aeruginosa sp. ZN was selected as high-efficiency biosurfactant producing strain from soil contaminated with oil from Ahvaz City, Khuzestan Province, southern Iran. The biosurfactant production in modified BH2 culture medium supplemented with 1% n-hexadecane occurred during exponential phase resulting in a reduction surface tension from 70 to 29 mN/m. Strain ZN produced biosurfactant with different properties to other Pseudomonas reported. These characterizations included continued production at C/N ratio range of 10-40; the produced biosurfactant could not separate stable emulsion of span-80-kerosene: Tween-80-distilled water (30:70) within 24 h. The produced biosurfactants were able to increase hydrophobicity of bacterial cell to 55%. Recovery of biosurfactants from cell-free supernatant was performed with acid precipitation and ammonium sulfate precipitation. Chemical analysis such as spraying techniques on developed TLC plate and staining methods of supernatant indicated that produced biosurfactants were glycolipids, characterized by ESI-MS analysis of extracted product as di-rhamnolipids.

Conclusion: Ability of this strain to produce biosurfactant in the presence of cooked oil and n-hexadecane make it an optimistic candidate for biodegradation of some derivatives of crude oil and food industry.

 

Franzetti A, Bestetti B, Caredda P et al (2008). Surface-active compounds and their role in the access to hydrocarbons in Gordonia strains. FEMS Microbiol Ecol, 63(2):238-48.

Sekhon K, Khanna S, Cameotra S (2011). Enhanced biosurfactant production through cloning of three genes and role of esterase in biosurfactant release. Microb Cell Fact, 10:49.

Pacwa-Płociniczak M, Płaza GA, Piotrowska-Seget Z, Cameotra SS (2011). Environmental applications of biosurfactants: recent advances. Int J Mol Sci, 12(1):633-54.

Banat I, Makkar R, Cameotra S (2000). Potential commercial applications of microbial surfactants. Appl Microbiol Biotechnol, 53(5):495-508.

Lotfabad T, Shourian M, Roostaazad R et al (2009). An efficient biosurfactant-producing bacterium Pseudomonas aeruginosa MR01, isolated from oil excavation areas in south of Iran. Colloids Surf B Biointerfaces, 69(2):183-93.

Abouseoud M, Maachi R, Amrane A et al (2008). Evaluation of different carbon and nitrogen sources in production of biosurfactant by Pseudomonas fluorescens. Desalination, 223: 143-51.

Nadarajah N, Singh A, Ward O (2002). Evaluation of a mixed bacterial culture for de-emulsification of water-in-petroleum oil emulsions. World J Microbiol Biotechnol, 18 (5): 435-40.

Youssef N, Duncan K, Nagle D et al (2004). Comparison of methods to detect biosurfactant production by diverse microorganisms. J Microbiol Methods, 56 (3): 339-47.

Biniarz P, Łukaszewicz M (2017). Direct quantification of lipopeptide biosurfactants in biological samples via HPLC and UPLC-MS requires sample modification with an organic solvent. Appl Microbiol Biotechnol, 101 (11): 4747-59.

Karthik L, Kumar G, Bhaskara Rao K (2010). Comparison of methods and screening of biosurfactant producing marine Actinobacteria isolated from Nicobar marine sediment. IIOAB Journal, 1 (2): 34-8.

Gudiña E, Fernandes E, Rodrigues A et al (2015). Biosurfactant production by Bacillus subtilis using corn steep liquor as culture medium. Front Microbiol, 6:59.

Govindammal M, Parthasarathi R (2013). Investigation on antimicrobial activity of biosurfactant produced by Pseudomonas fluorescens isolated from mangrove ecosystem. Int Res J Pharmacy, 4 (1): 230-32.

Abalos A, Pinazo A, Infante M et al (2001). Physicochemical and Antimicrobial Properties of New Rhamnolipids Produced by Pseudomonas aeruginosa AT10 from Soybean Oil Refinery Wastes. Langmuir, 17 (5): 1367-71.

Singh V (2012). Biosurfactant–Isolation, production, purification & significance. Int J Sci Res Pub, 2 (7): 1-4.

Smyth T, Perfumo A, Marchant R, Banat I (2010). Isolation and analysis of low molecular weight microbial glycolipids. In: Handbook of hydrocarbon and lipid microbiology. Ed,Timmis. Springer-verlag Berlin Heidelberg, pp. 3705-3723.

Smyth TJP, Perfumo A, McClean S et al (2010). Isolation and Analysis of Lipopeptides and High Molecular Weight Biosurfactants. In: Handbook of Hydrocarbon and Lipid Microbiology. Ed,Timmis. Springer-verlag Berlin Heidelberg, pp. 3687-3704.

Saikia R, Deka S, Deka M, Banat I (2012). Isolation of biosurfactant-producing Pseudomonas aeruginosa RS29 from oil-contaminated soil and evaluation of different nitrogen sources in biosurfactant production. Ann Microbiol, 62 (2): 753-63.

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

Herman D, Maier R (2002). Biosynthesis and applications of glycolipid and lipopeptide biosurfactants. In: Lipid Biotechnology. Eds, Kuo and Gardner. Marcel Dekkar, Inc. New York, pp. 629-654.

Hamzah A, Sabturani N, Radiman S (2013). Screening and Optimization of Biosurfactant Production by the Hydrocarbon-degrading Bacteria. Sains Malays, 42 (5): 615-23.

Raza ZA, Ahmad N, Kamal S (2014). Multi-response optimization of rhamnolipid production using grey rational analysis in Taguchi method. Biotechnol Rep (Amst), 3: 86-94.

Zhao Z, Selvam A, Wong JWC (2011). Effects of rhamnolipids on cell surface hydrophobicity of PAH degrading bacteria and the biodegradation of phenanthrene. Bioresour Technol, 102 (5): 3999-4007.

Meng Q (2015). Application of rhamnolipid as demulsifier. Patent No. CN 102851059 B. https://patents.google.com/patent/CN102851059B/en?oq=Application+of+rhamnolipid+as+demulsifier.++Patent+No.+CN+102851059+B

Long X, Zhang G, Shen C et al (2013). Application of rhamnolipid as a novel biodemulsifier for destabilizing waste crude oil. Bioresour Technol, 131:1-5.

Coutinho JOPA, Silva MPS, Moraes PM et al (2013). Demulsifying properties of extracellular products and cells of Pseudomonas aeruginosa MSJ isolated from petroleum-contaminated soil. Bioresour Technol, 128: 646-54.

Mohammed R, Bailey A, Luckham P, Taylor S (1994). Dewatering of crude oil emulsions 3. Emulsion resolution by chemical means. Colloids Surf A Physicochem Eng Asp, 83 (3): 261-71.

Ly MH, Aguedo M, Goudot S et al (2008). Interactions between bacterial surfaces and milk proteins, impact on food emulsions stability. Food Hydrocoll, 22 (5): 742-51.

Janek T, Łukaszewicz M, Rezanka T, Krasowska A (2010). Isolation and characterization of two new lipopeptide biosurfactants produced by Pseudomonas fluorescens BD5 isolated from water from the Arctic Archipelago of Svalbard. Bioresour Technol, 101 (15): 6118-23.

Kuiper I, Lagendijk E, Pickford R et al (2004). Characterization of two Pseudomonas putida lipopeptide biosurfactants, putisolvin I and II, which inhibit biofilm formation and break down existing biofilms. Mol Microbiol, 51 (1): 97-113.

Smyth T, Rudden M, Tsaousi K et al (2014). Protocols for the Isolation and Analysis of Lipopeptides and Bioemulsifiers. In: Hydrocarbon and Lipid Microbiology Protocols, Springer Protocols Handbooks. Ed, McGenity. Springer-Verlag Berlin Heidelberg, pp. 3-28.

Smyth T, Rudden M, Tsaousi K et al (2014). Protocols for the Detection and Chemical Characterisation of Microbial Glycolipids. In: Hydrocarbon and Lipid Microbiology Protocols, Springer Protocols Handbooks. Ed, McGenity. Springer-Verlag Berlin Heidelberg, pp. 29-60.

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IssueVol 47 No 8 (2018) QRcode
SectionOriginal Article(s)
Keywords
Screening Pseudomonas aeruginosa Characterization Rhamnolipids

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How to Cite
1.
NAJMI Z, EBRAHIMIPOUR G, FRANZETTI A, BANAT IM. Investigation of Physicho-chemical Properties and Characterization of Produced Biosurfactant by Selected Indigenous Oil-degrading Bacterium. Iran J Public Health. 2018;47(8):1151-1159.