Environmental Science and Pollution Research

, Volume 24, Issue 22, pp 18699–18709 | Cite as

Soy molasses as a fermentation substrate for the production of biosurfactant using Pseudomonas aeruginosa ATCC 10145

  • Marília Silva Rodrigues
  • Felipe Santos Moreira
  • Vicelma Luiz Cardoso
  • Miriam Maria de ResendeEmail author
Short Research and Discussion Article


Soy molasses is a product co-generated during soybean processing that has high production and low commercial value. Its use has great potential in fermentative processes due to the high concentration of carbohydrates, lipids and proteins. This study investigated the use of Pseudomonas aeruginosa to produce biosurfactants in a soy molasses-based fermentation medium. A central composite design (CCD) was prepared with two variables and three replicates at the central point to optimize the production of biosurfactant. The concentration of soy molasses had values between 29.3 and 170.7 g/L and the initial concentration of microorganism varied between 0.2 and 5.8 g/L. All the experiments were performed in duplicate on a shaker table at 30.0 ± 1.0 °C and 120 rpm for 72 h with samples taken every 12 h. Thus, to validate the experiments, the values of 120 g/L for the initial concentration of soy molasses and 4 g/L for the initial concentration of microorganisms were used. In response, the following values were obtained at 48 h of fermentation: surface tension of 31.9 dyne/cm, emulsifying index of 97.4%, biomass concentration of 11.5 g/L, rhamnose concentration of 6.9 g/L and biosurfactant concentration of 11.70 g/L. Further analysis was carried out for critical micelle concentration (CMC) which was obtained at approximately 80 mg/L. The bands found in Fourier transform infrared spectroscopy analysis had characteristic glycolipids as reported in the literature. These values show a great potential for biosurfactant production using soy molasses as a substrate and bacteria of the species P. aeruginosa.


Use of agro-industrial waste Value-aggregated bioproduct Environmentally friendly process 



This work was supported by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), the National Council for Technological and Scientific Development (CNPq) and Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), Brazil.


  1. Abouseoud M, Maachi R, Amrane A, Boudergua S, Nabi A (2008) Evaluation of different carbon and nitrogen sources in production of biosurfactant by Pseudomonas fluorescens. Desalination 223:143–151CrossRefGoogle Scholar
  2. Abyaneh AS, Fazaelipoor MH (2016) Evaluation of rhamnolipid (RL) as a biosurfactant for the removal of chromium from aqueous solution by precipitate flotation. J Environ Management 165:184–187CrossRefGoogle Scholar
  3. Arino S, Marchal R, Vandecasteele JP (1996) Identification and production of a rhamnolipidic biosurfactant by a Pseudomonas species. Appl Microbiol Biotechnol 45(162):168Google Scholar
  4. Banat IM, Franzetti A, Gandolfi I, Bestetti G, Marinotti MG, Fracchia L, Smyth TJ, Marchant R (2010) Microbial biosurfactants production, applications and future potential. Appl Microbiol Biotechnol 87:427–444CrossRefGoogle Scholar
  5. Benincasa M, Contiero J, Manresa MA, Moraes IO (2002) Rhamnolipid production by Pseudomonas aeruginosa LBI growing on soapstock as the carbon source. J Food Eng Essex 54:283–288CrossRefGoogle Scholar
  6. 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. Anton Leeuw, Amsterdam 85:1–8CrossRefGoogle Scholar
  7. da Borges WS, Cardoso VL, de Resende MM (2012) Use of a greasy effluent floater treatment station from the slaughterhouse for biosurfactant production. Biotechnol Appl Bioc 59:238–244CrossRefGoogle Scholar
  8. Borges WS, Moura AAO, Coutinho Filho U, Cardoso VL, Resende M (2015) Optimization of the operating conditions for rhamnolipid production using slaughterhouse-generated industrial float as substrate. Braz J Chem Eng 32:357–365CrossRefGoogle Scholar
  9. Cooper DG, Goldenbenberg BG (1987) Surface-active agents from two Bacillus species. Appl Environ Microbiol 42:224–229Google Scholar
  10. Deb M, Mandal N, Sathiavelu M, Arunachalam S (2016) Application and future aspects of microbial biosurfactants – review. Res J Pharm Biol Chem Sci 7:2803–2812Google Scholar
  11. Deepika KV, Sridhar PR, Bramhachari PV (2015) Characterization and antifungal properties of rhamnolipids produced by mangrove sediment bacterium Pseudomonas aeruginosa strain KVD-HM52. Biocatal Agr Biotechnol 4:608–615Google Scholar
  12. Deepika KV, Kalam S, Sridhar PR, Podile AR, Bramhachari PV (2016) Optimization of rhamnolipid biosurfactant production by mangrove sediment bacterium Pseudomonas aeruginosa KVD-HR42 using response surface methodology. Biocatal Agr Biotechnol 5:38–47Google Scholar
  13. Dwivedi S, Saquib Q, Al-Khedhairy AA, Ahmad J, Siddiqui MA, Musarrat J (2015) Rhamnolipids functionalized AgNPs-induced oxidative stress and modulation of toxicity pathway genes in cultures MCF-7 cells. Colloid Surface B 132:290–298CrossRefGoogle Scholar
  14. Guerra-Santos L, Kappeli O, Fiechter A (1984) Pseudomonas aeruginosa biosurfactant production in continuous culture with glucose as carbon source. Appl Environ Microbiol 48:301–305Google Scholar
  15. Haba E, Espuny MJ, Busquets M, Manresa A (2000) Screening and production of rhamnolipids by Pseudomonas aeruginosa 47 T2 NCIB 40044 from waste frying oils. J Appl Microbiol, Oxford 88:379–387CrossRefGoogle Scholar
  16. Johnson LA, Myers DJ, Burden DJ (1992) Soy protein’s history, prospects in food and feed. Inform 3:429–444Google Scholar
  17. Joy S, Rahma, PKSM, Sharma S (2017) Biosurfactant production and concomitant hydrocarbon degradation potentials of bacteria isolated from extreme and hydrocarbon contaminated environments. Cheml Eng J 317:232–241Google Scholar
  18. Lan G, Fan Q, Liu Y, Chen C, Li G, Liu Y, Yin X (2015) Rhamnolipid production from waste cooking oil using Pseudomonas SWP-4. Biochem Eng J 101:44–54CrossRefGoogle Scholar
  19. Lang S (2002) Biological amphiphiles: microbial surfactants. Curr Opin Interf Sci 74:59–70Google Scholar
  20. de Lima CJB, França FP, Sérvulo EFC, de Resende MM, Cardoso VL (2007) Enhancement of rhamnolipid production in residual soybean oil by an isolated strain of Pseudomonas aeruginosa. Appl Biochem Biotechnol 137:463–470Google Scholar
  21. de Lima CJB, Ribeiro EJ, Sérvulo EFC, de Resende MM, Cardoso VL (2009) Biosurfactant production by Pseudomonas aeruginosa grown in residual soybean oil. Appl Biochem Biotechnol 152:156–168CrossRefGoogle Scholar
  22. Ma KY, Sun MY, Dong W, He CQ, Chen FL, Ma YL (2016) Effects of nutrition optimization strategy on rhamnolipid production in a Pseudomonas aeruginosa strain DN1 for bioremediation of crude oil. Biocatal Agr Biotechol 6:144–151Google Scholar
  23. Mao X, Jiang R, Xiao W, Yu J (2015) Use of surfactants for the remediation of contaminated soils: a review. J Hazard Mater 285:419–435CrossRefGoogle Scholar
  24. Massara H, Mulligan CN, Hadjinicolaou J (2007) Effect of rhamnolipids on chromium-contaminated kaolinite. Soil Sediment Contam 16:1–14CrossRefGoogle Scholar
  25. Monteiro AS (2007) Caracterização Molecular e Estrutural de Biosurfactantes Produzidos por Pseudomonas aeruginosa UFPEDA 614. (Tese). UFPR, 2007. Disponível em:;jsessionid=02600E587273588D38A30761AEE971FA?sequence=1.
  26. Morillo E, Villaverde J (2017) Advanced technologies for the remediation of pesticide-contaminated soils. Sci Total Environ 586:576–597CrossRefGoogle Scholar
  27. Mulligan CN (2005) Environmental applications for biosurfactants. Environ Pollut 133:183–198Google Scholar
  28. Mulligan C (2009) Recent advances in the environmental applications of biosurfactants. Colloid Interf Sci 14:372–378CrossRefGoogle Scholar
  29. Mulligan CN, Gibbs BF (1989) Correlation of nitrogen metabolism with biosurfactant production by Pseudomonas aeruginosa. Appl Environ Microbiol 55:3016–3019Google Scholar
  30. Nistchke M, Pastore GM (2002) Biossurfactantes: Propriedades e Aplicações. Rev Quim Nova 25:772–776CrossRefGoogle Scholar
  31. Plociniczak MP, Plaza GA, Seget ZP, Cameotra SS (2011) Environmental applications of biosurfactants: recent advances. Int J Mol Sci 12:633–654CrossRefGoogle Scholar
  32. Prabakaran G, Hoti SL, Rao HSP, Vijjapu S (2015) Di-rhamnolipid is a mosquito pupicidal metabolite from Pseudomonas fluorescens (VCRC B426). Acta Trop 148:24–31CrossRefGoogle Scholar
  33. Qureshi N, Lolas A, Blaschek HP (2001) Soy molasses as fermentation substrate for production of butanol using Clostridium beijerinckii BA 101. J Ind Microbiol Biotechnol 26:290–295CrossRefGoogle Scholar
  34. Rahman KSM, Rahman TJ, Mcclean S, Marchant R, Banat IM (2002) Rhamnolipid biosurfactant production by strains of Pseudomonas aeruginosa using low-cost raw materials. Biotechnol Prog 18:1277–1281CrossRefGoogle Scholar
  35. Ramiréz IM, Tsaousi K, Rudden M, Marchant R, Alameda EJ, Román MG, Banat IM (2015) Rhamnolipid and surfactin production from olive oil mill waste as sole carbon source. Bioresour Technol 198:231–236CrossRefGoogle Scholar
  36. Raza ZA, Rehman A, Khan MS, Khalid ZM (2007) Improved production of biosurfactant by a Pseudomonas aeruginosa mutant using vegetable oil refinery wastes. Biodegradation 18:115–121CrossRefGoogle Scholar
  37. Raza ZA, Rheman A, Hussain MT, Masood R, Haq A, Saddique MT, Javid A, Ahmad N (2014) Production of rhamnolipid surfactant and its application in bioscouring of cotton fabric. Carbohydr Res 391:97–105CrossRefGoogle Scholar
  38. Romão BB, da Silva FB, Cardoso VL, de Resende MM (2012) Ethanol production from hydrolyzed soybean molasses. Energy & Fuels (Print) 26:2310–2316CrossRefGoogle Scholar
  39. Santana Filho AP (2009) Ramnolipídeos produzidos por Pseudomonas aeruginosa UFPEDA 614: estudos de produção e de variação da composição de homólogos. (Dissertação) UFP. Disponível em:ção%20arquimedes%20p%20santana%20filho.pdf?sequence=1
  40. Santos Lopes V, Fischer J, Pinheiro TMA, Cabral BV, Cardoso VL, Coutinho Filho U (2017) Biosurfactant and ethanol co-production using Pseudomonas aeruginosa and Saccharomyces cerevisiae co-cultures and exploded sugarcane bagasse. Renew Energy 109:305–310CrossRefGoogle Scholar
  41. Siqueira PF (2007) Production of bio-ethanol from soybean molasses by Saccharomyces cerevisiae. Master Dissertation, Federal University of Paraná/Universities of Provence and of the Mediterranean.Google Scholar
  42. Smith AK, Circle SJ (1978) Historical background. Soybeans: chemistry and technology. v. 1, Westport: The AVI publishing company, pp. 1–26Google Scholar
  43. Torres LG, Ramos F, Avila MA, Ortiz I (2012) Removal of methyl parathion by surfactant-assisted soil washing and subsequent wastewater biological treatment. J Pestic Sci 37:240–246CrossRefGoogle Scholar
  44. Wan J, Meng D, Long T, Ying R, Ye M, Zhang S, Li Q, Zhou Y, Lin Y (2015) Simultaneous removal of lindane, lead and cadmium from soils by rhamnolipids combined with citric acid. PLoS One 10(6):e0129978. doi: 10.1371/journal.pone.0129978 CrossRefGoogle Scholar
  45. Wu Z, Zhong H, Yuan X, Wang H, Wang L, Chen X, Zeng G, Wu Y (2014) Adsorptive removal of methylene blue by rhamnolipid-functionalized graphene oxide from wastewater. Water Res 67:330–344CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Marília Silva Rodrigues
    • 1
  • Felipe Santos Moreira
    • 1
  • Vicelma Luiz Cardoso
    • 1
  • Miriam Maria de Resende
    • 1
    Email author
  1. 1.Chemical Engineering FacultyFederal University of UberlândiaUberlândiaBrazil

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