Characterization and properties of the biosurfactant produced by PAH-degrading bacteria isolated from contaminated oily sludge environment

  • Varsha Tripathi
  • Vivek Kumar Gaur
  • Nitesh Dhiman
  • Krishna Gautam
  • Natesan ManickamEmail author
Sustainable Industrial and Environmental Bioprocesses


The aim of the present study was to investigate biosurfactant production ability of five different polyaromatic hydrocarbon (PAH)-metabolizing bacteria, such as Ochrobactrum anthropi IITR07, Pseudomonas mendocina IITR46, Microbacterium esteraromaticum IITR47, Pseudomonas aeruginosa IITR48, and Stenotrophomonas maltophilia IITR87. These bacteria showed biosurfactant production using 2% glucose as rich substrate; strain IITR47 yielded the highest with 906 and 534 mg/L biosurfactant in the presence of naphthalene and crude oil as the unique carbon sources. P. aeruginosa IITR48 showed the least surface tension at 29 N/m and the highest emulsification index at 63%. The biosurfactants produced were identified as glycolipid and rhamnolipid based on Fourier transform infrared spectroscopy analysis. In particular, the biosurfactant produced by bacteria S. maltophilia IITR87 efficiently emulsified mustard oil with an E24 value of 56%. It was observed that, all five biosurfactants from these degrader strains removed 2.4-, 1.7-, 0.9-, 3.8-, and 8.3-fold, respectively, crude oil from contaminated cotton cloth. Rhamnolipid derived from IITR87 was most efficient, exhibiting highest desorption of crude oil. These biosurfactants exhibited good stability without significantly losing its emulsification ability under extreme conditions, thus can be employed for bioremediation of PAHs from diverse contaminated ecosystem.

Graphical Abstract


Biosurfactant Biodegradation Emulsification Crude oil Rhamnolipid Antibacterial 



VT is thankful to UP-CST for providing fellowship. VKG is thankful to Council of Scientific and Industrial Research (CSIR), India for providing senior research fellowship. This manuscript bears IITR communication number 3580.

Funding information

This study was financially supported by the UP Council of Science and Technology (UP-CST), Uttar Pradesh, India, under the project GAP-360.


  1. Benincasa M, Abalos A, Oliveira I, Manresa A (2004) Chemical structure, surface properties and biological activities of the biosurfactant produced by Pseudomonas aeruginosa LBI from soapstock. Antonie Van Leeuwenhoek 85:1–8CrossRefGoogle Scholar
  2. Bouassida M, Fourati N, Ghazala I, Ellouze-Chaabouni S, Ghribi D (2018) Potential application of Bacillus subtilis SPB1 biosurfactants in laundry detergent formulations: Compatibility study with detergent ingredients and washing performance. Eng Life Sci 18:70–77CrossRefGoogle Scholar
  3. Cai Q, Zhang B, Chen B, Zhu Z, Lin W, Cao T (2014) Screening of biosurfactant producers from petroleum hydrocarbon contaminated sources in cold marine environments. Mar Pollut Bull 86:402–410CrossRefGoogle Scholar
  4. Campos JM, Stamford TL, Rufino RD, Luna JM, Stamford TC, Sarubbo LA (2015) Formulation of mayonnaise with the addition of a bioemulsifier isolated from Candida utilis. Toxicol Rep 2:1164–1170CrossRefGoogle Scholar
  5. Chebbi A, Hentati D, Cheffi M, Bouabdallah R, Choura C, Sayadi S, Chamkha M (2018) Promising abilities of mercapto-degrading Staphylococcus capitis strain SH6 in both crude oil and waste motor oil as sole carbon and energy sources: its biosurfactant production and preliminary characterization. J Chem Technol Biotechnol 93:1401–1412CrossRefGoogle Scholar
  6. Chen Q, Bao M, Fan X, Liang S, Sun P (2013) Rhamnolipids enhance marine oil spill bioremediation in laboratory system. Mar Pollut Bull 71:269–275CrossRefGoogle Scholar
  7. Chen C, Sun N, Li D, Long S, Tang X, Xiao G, Wang L (2018) Optimization and characterization of biosurfactant production from kitchen waste oil using Pseudomonas aeruginosa. Environ Sci Pollut Res 25:14934–14943CrossRefGoogle Scholar
  8. Das P, Mukherjee S, Sen R (2008) Antimicrobial potential of a lipopeptide biosurfactant derived from a marine Bacillus circulans. J Appl Microbiol 104:1675–1684CrossRefGoogle Scholar
  9. Datta P, Tiwari P, Pandey LM (2018) Isolation and characterization of biosurfactant producing and oil degrading Bacillus subtilis MG495086 from formation water of Assam oil reservoir and its suitability for enhanced oil recovery. Bioresour Technol 270:439–448CrossRefGoogle Scholar
  10. Gargouri B, del Mar Contreras M, Ammar S, Segura-Carretero A, Bouaziz M (2017) Biosurfactant production by the crude oil degrading Stenotrophomonas sp. B-2: chemical characterization, biological activities and environmental applications. Environ Sci Pollut Res 24:3769–3779CrossRefGoogle Scholar
  11. Gaubert A, Clement Y, Bonhomme A, Burger B, Bouveresse DJ, Rutledge D, Casabianca H, Lanteri P, Bordes C (2016) Characterization of surfactant complex mixtures using Raman spectroscopy and signal extraction methods: application to laundry detergent deformulation. Anal Chim Acta 915:36–48CrossRefGoogle Scholar
  12. Gaur VK, Bajaj A, Regar RK, Kamthan M, Jha RR, Srivastava JK, Manickam N (2019) Rhamnolipid from a Lysinibacillus sphaericus strain IITR51 and its potential application for dissolution of hydrophobic pesticides. Bioresour Technol 272:19–25CrossRefGoogle Scholar
  13. Jain RM, Mody K, Mishra A, Jha B (2012) Physicochemical characterization of biosurfactant and its potential to remove oil from soil and cotton cloth. Carbohydr Polym 89:1110–1116CrossRefGoogle Scholar
  14. Jimoh AA, Lin J (2018) Enhancement of Paenibacillus sp. D9 Lipopeptide biosurfactant production through the optimization of medium composition and its application for biodegradation of hydrophobic pollutants. Appl Biochem Biotechnol 25:1–20Google Scholar
  15. Johnsen AR, Wick LY, Harms H (2005) Principles of microbial PAH-degradation in soil. Environ Pollut 133:71–84CrossRefGoogle Scholar
  16. Joy S, Butalia T, Sharma S, Rahman PKSM (2017) Biosurfactant producing bacteria from hydrocarbon contaminated environment. In: Heimann K., Karthikeyan O., Muthu S. (eds) Biodegradation and bioconversion of hydrocarbons. Environmental footprints and eco-design of products and processes. Springer, Singapore, 259-305Google Scholar
  17. Kalyani ALT, Sireesha GN, Sankar GGG, Prabhakar T (2014) Isolation of bio-surfactant producing actinomycetes from terrestrial and marine soils. Int J Pharm Sci Res 5:4015Google Scholar
  18. Karlapudi AP, Venkateswarulu TC, Tammineedi J, Kanumuri L, Ravuru BK, ramu Dirisala V, Kodali VP (2018) Role of biosurfactants in bioremediation of oil pollution-a review. Petroleum 4:241–249CrossRefGoogle Scholar
  19. Khalid HF, Tehseen B, Sarwar Y, Hussain SZ, Khan WS, Raza ZA, Bajwa SZ, Kanaras AG, Hussain I, Rehman A (2019) Biosurfactant coated silver and iron oxide nanoparticles with enhanced anti-biofilm and anti-adhesive properties. J Hazard Mater 364:441–448CrossRefGoogle Scholar
  20. Khopade A, Ren B, Liu XY, Mahadik K, Zhang L, Kokare C (2012) Production and characterization of biosurfactant from marine Streptomyces species B3. J Colloid Interface Sci 367:311–318CrossRefGoogle Scholar
  21. Kumar CG, Sujitha P, Mamidyala SK, Usharani P, Das B, Reddy CR (2014) Ochrosin, a new biosurfactant produced by halophilic Ochrobactrum sp. strain BS-206 (MTCC 5720): purification, characterization and its biological evaluation. Process Biochem 49:1708–1717CrossRefGoogle Scholar
  22. Kumari S, Regar RK, Bajaj A, Ch R, Satyanarayana GN, Mudiam MK, Manickam N (2017) Simultaneous biodegradation of polyaromatic hydrocarbons by a Stenotrophomonas sp: characterization of nid genes and effect of surfactants on degradation. Indian J Microbiol 57:60–67CrossRefGoogle Scholar
  23. Kumari S, Regar RK, Manickam N (2018) Improved polycyclic aromatic hydrocarbon degradation in a crude oil by individual and a consortium of bacteria. Bioresour Technol 254:174–179CrossRefGoogle Scholar
  24. Li B, Cai D, Hu S, Zhu A, He Z, Chen S (2018) Enhanced synthesis of poly gamma glutamic acid by increasing the intracellular reactive oxygen species in the Bacillus licheniformis Δ1-pyrroline-5-carboxylate dehydrogenase gene ycgN-deficient strain. Appl Microbiol Biotechnol 102:10127–10137CrossRefGoogle Scholar
  25. Manickam N, Bajaj A, Saini HS, Shanker R (2012) Surfactant mediated enhanced biodegradation of hexachlorocyclohexane (HCH) isomers by Sphingomonas sp. NM05. Biodegradation 23:673–682CrossRefGoogle Scholar
  26. Mesbaiah FZ, Eddouaouda K, Badis A, Chebbi A, Hentati D, Sayadi S, Chamkha M (2016) Preliminary characterization of biosurfactant produced by a PAH-degrading Paenibacillus sp. under thermophilic conditions. Environ Sci Pollut Res 23:14221–14230CrossRefGoogle Scholar
  27. Mouafo TH, Mbawala A, Ndjouenkeu R (2018) Effect of different carbon sources on biosurfactants production by three strains of Lactobacillus spp.. BioMed Research International Article ID. 5034783 vol. 2018:15. Scholar
  28. Nitschke M, Pastore GM (2004) Biosurfactant production by Bacillus subtilis using cassava-processing effluent. Appl Biochem Biotechnol 112(3):163–172Google Scholar
  29. Pei G, Sun C, Zhu Y, Shi W, Li H (2018) Biosurfactant-enhanced removal of o, p-dichlorobenzene from contaminated soil. Environ Sci Pollut Res 25:18–26CrossRefGoogle Scholar
  30. Pereira JF, Gudiña EJ, Costa R, Vitorino R, Teixeira JA, Coutinho JA, Rodrigues LR (2013) Optimization and characterization of biosurfactant production by Bacillus subtilis isolates towards microbial enhanced oil recovery applications. Fuel 111:259–268CrossRefGoogle Scholar
  31. Pornsunthorntawee O, Wongpanit P, Chavadej S, Abe M, Rujiravanit R (2008) Structural and physicochemical characterization of crude biosurfactant produced by Pseudomonas aeruginosa SP4 isolated from petroleum-contaminated soil. Bioresour Technol 99:1589–1595CrossRefGoogle Scholar
  32. Rodríguez-Lopez L, Rincón-Fontán M, Vecino X, Cruz JM, Moldes A (2017) Ionic behavior assessment of surface-active compounds from corn steep liquor by exchange resins. J Surfactant Deterg 20:207–217CrossRefGoogle Scholar
  33. Satpute SK, Banat IM, Dhakephalkar PK, Banpurkar AG, Chopade BA (2010) Biosurfactants, bioemulsifiers and exopolysaccharides from marine microorganisms. Biotechnol Adv 28:436–450CrossRefGoogle Scholar
  34. Sharma D, Ansari MJ, Al-Ghamdi A, Adgaba N, Khan KA, Pruthi V, Al-Waili N (2015) Biosurfactant production by Pseudomonas aeruginosa DSVP20 isolated from petroleum hydrocarbon-contaminated soil and its physicochemical characterization. Environ Sci Pollut Res 22:17636–17643CrossRefGoogle Scholar
  35. Silva SN, Farias CB, Rufino RD, Luna JM, Sarubbo LA (2010) Glycerol as substrate for the production of biosurfactant by Pseudomonas aeruginosa UCP0992. Colloids Surf B: Biointerfaces 79:174–183CrossRefGoogle Scholar
  36. Singh AK, Cameotra SS (2013) Efficiency of lipopeptide biosurfactants in removal of petroleum hydrocarbons and heavy metals from contaminated soil. Environ Sci Pollut Res 20:7367–7376CrossRefGoogle Scholar
  37. Stallwood B, Shears J, Williams PA, Hughes KA (2005) Low temperature bioremediation of oil-contaminated soil using biostimulation and bioaugmentation with a Pseudomonas sp. from maritime Antarctica. J Appl Microbiol 99:794–802CrossRefGoogle Scholar
  38. Thampayak I, Cheeptham N, Pathom-Aree W, Leelapornpisid P, Lumyong S (2008) Isolation and identification of biosurfactant producing Actinomycetes from soil. Res J Microbiol 3:499–507CrossRefGoogle Scholar
  39. Thavasi R, Sharma S, Jayalakshmi S (2011) Evaluation of screening methods for the isolation of biosurfactant producing marine bacteria. J Pet Environ Biotechnol 1:1–6Google Scholar
  40. Tichy J, Novak J (1998) Extraction, assay, and analysis of antimicrobials from plants with activity against dental pathogens (Streptococcus sp.). J Altern Complement Med 4:39–45CrossRefGoogle Scholar
  41. Tuleva B, Christova N, Jordanov B, Nikolova-Damyanova B, Petrov P (2005) Naphthalene degradation and biosurfactant activity by Bacillus cereus 28BN. Z Naturforsch 60:577–582CrossRefGoogle Scholar
  42. Wadekar SD, Kale SB, Lali AM, Bhowmick DN, Pratap AP (2012) Microbial synthesis of rhamnolipids by Pseudomonas aeruginosa (ATCC 10145) on waste frying oil as low cost carbon source. Preparative Biochem Biotechnol 42(3):249–266Google Scholar
  43. Wittgens A, Tiso T, Arndt TT, Wenk P, Hemmerich J, Müller C, Wichmann R, Küpper B, Zwick M, Wilhelm S, Hausmann R (2011) Growth independent rhamnolipid production from glucose using the non-pathogenic Pseudomonas putida KT2440. Microb Cell Factories 10:8CrossRefGoogle Scholar
  44. Xia W, Zhi-Bin L, Han-Ping D, Li Y, Qing-Feng C, Yong-Qiang (2012) Biosynthesis, characterization, and oil recovery application of biosurfactant produced by indigenous Pseudomonas aeruginosa WJ-1 using waste vegetable oils. Appl Biochem Biotechnol 166(5):1148–1166Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Varsha Tripathi
    • 1
  • Vivek Kumar Gaur
    • 1
    • 2
  • Nitesh Dhiman
    • 3
    • 4
  • Krishna Gautam
    • 1
    • 4
  • Natesan Manickam
    • 1
    Email author
  1. 1.Environmental Biotechnology Division, Environmental Toxicology GroupCSIR-Indian Institute of Toxicology ResearchLucknowIndia
  2. 2.Amity Institute of BiotechnologyAmity University Uttar Pradesh, Lucknow CampusLucknow 226010India
  3. 3.Water Analysis Laboratory, Nanomaterial Toxicology GroupCSIR-Indian Institute of Toxicology ResearchLucknowIndia
  4. 4.Academy of Scientific and Innovative Research (AcSIR)CSIR-Indian Institute of Toxicology ResearchLucknowIndia

Personalised recommendations