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Frankfurt am Main-BRD , 2015

SURFACE ACTIVITY OF SURFACTIN UNDER EXTREME CONDITIONS ( p H, TEMPERATURE AND IONIC STRENGTH) – A PROSPECTION FOR MICROBIAL ENHANCED OIL RECOVERY. PhD student – Cristiano J. Andrade Supervisior – Dr. Glaucia M. Pastore Co-supervisior – Dr. Francisco F. C. Barros

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Frankfurt am Main-BRD , 2015

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  1. SURFACE ACTIVITY OF SURFACTIN UNDER EXTREME CONDITIONS (pH, TEMPERATURE AND IONIC STRENGTH) – A PROSPECTION FOR MICROBIAL ENHANCED OIL RECOVERY PhD student – Cristiano J. Andrade Supervisior – Dr. Glaucia M. Pastore Co-supervisior – Dr. Francisco F. C. Barros Supervisior (Split PhD) – Dr. Paula Jauregi Frankfurt amMain-BRD, 2015

  2. THE AIM OF THIS STUDY WAS... • Biotechnologicalproduction • Purification • Chemicalstructureidentification • Application - MEOR

  3. SCREENING OF BIOSURFACTANT PRODUCERS PRODUCTION IDENTIFICATION PURIFICATION APPLICATION Bacillus subtilis LB5a

  4. SCREENING OF BIOSURFACTANT PRODUCERS PRODUCTION IDENTIFICATION PURIFICATION APPLICATION SURFACTIN – lipopeptide Bacillus subtilis LB5a

  5. PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  6. COMPOSITION OF CULTURE MEDIUM PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  7. PRODUCTION STRATEGIES PRODUCTION IDENTIFICATION Continuous fermentation Batch fermentation Feed-Batch fermentation Flask (250 mL) Top-bench bioreactor (7 L) Pilot scale (80 L) Top-bench bioreactor (7 L) Top-bench bioreactor (7 L) PURIFICATION APPLICATION Simultaneous production diacetyl (2,3-butanedione), acetoin (3-hydroxy-2-butanone), and 2,3-butanediol, Proteases, amylases and lipases

  8. PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  9. PUBLICATIONS PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  10. PURIFICATION…BUT HOW TO DO IT? PRODUCTION IDENTIFICATION PURIFICATION APPLICATION Ultrafiltration in two steps of surfactin produced by Bacillus subtilis LB5a using cassava wastewater as substrate

  11. PRODUCTION IDENTIFICATION Criticalmicelleconcentration (CMC) PURIFICATION APPLICATION • Surfactin - CMC ≈10 mg/L • Rhamnolipids- CMC ≈20 a 40 mg/L • Synthetic surfactants - CMC from 590 to 2000 mg/L

  12. PRODUCTION IDENTIFICATION Retentate Feed UF -1 Feed UF -2 Retentate UF-1 UF-2 PURIFICATION APPLICATION Permeate Low molecular weight

  13. PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  14. PRODUCTION IDENTIFICATION NMR PURIFICATION APPLICATION MALDI-TOFMS

  15. PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  16. PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  17. POTENTIAL APPLICATIONS PRODUCTION IDENTIFICATION • Antimicrobial • Emulsifier • Therapeuticagents • Pesticide • Food • Cosmetic • Etc PURIFICATION APPLICATION • MICROBIAL ENHANCE OIL RECOVERY

  18. MEOR PRODUCTION IDENTIFICATION Thus, the aim of microbial Enhanced Oil Recovery technologies is mainly the remaining oil in the wells – after the primary and secondary techniques - which is ≈3 trillion barrels (Shibulal et al. 2014 - Scientific World Journal). PURIFICATION APPLICATION PRIMARY RECOVERY SECONDARY RECOVERY

  19. MEOR PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  20. MEOR PRODUCTION IDENTIFICATION PURIFICATION APPLICATION • It was described that in the Daqingoilfield, the temperature ranged from 45 to 89oC and the ionic strength ≈15g.L-1, that is, the interaction of these parameters has to be consider.

  21. MEOR PRODUCTION IDENTIFICATION • This study describes for the first time, the surface activity measurements of surfactin at extreme conditions of temperature, ionic strength and pH (one at a time) and its interaction . PURIFICATION APPLICATION

  22. MEOR PRODUCTION IDENTIFICATION ONE A TIME PURIFICATION APPLICATION Temperature does notaffectthesurfaceactivityofsurfactin

  23. Central composite design gggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggg PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  24. MEOR PRODUCTION IDENTIFICATION PURIFICATION APPLICATION

  25. MEOR PRODUCTION IDENTIFICATION The ANOVA of ST and CMD-1 indicates that parameters (temperature, pH and ionic strength) are statistically different (95% of confidence) [(Fcalregression 23.02; Ftab 3.74); (Fcal lack of fit 5.01; Ftab19.41)] - (r2 of 0.76). Thus, the CCD analysis of surfactin - CMD-1 – resulted the following equation: Equation (1): Y = 67.28 + 8.5x – 3.02x2 Y is CMD-1 of surfactin, x ionic strength and x2 ionic strength squared. The ionic strength is the most significant parameters on the surface activity. It was observed that pH minimally influences the surface activity of surfactin. The same trend that temperature. The derivate of equation 1 with Y´=0 (maximum– red area) indicates that at 18.58 g.L-1 of brine is the threshold, when associate to extreme pH and temperature, in order to keep the surface activity. PURIFICATION APPLICATION

  26. MEOR PRODUCTION IDENTIFICATION heavy light medium PURIFICATION APPLICATION

  27. CONCLUSION • Thesurfactin, thatwasproducedby cassava wastewater as culturemediumandpurifiedbyultrafiltration, hasremarkablesurfaceactiviyunder extreme conditionsandalso great potential to beused for MEOR .

  28. eng.crisja@gmail.com Dankeschön

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