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Efficient production of 1,3-propanediol from fermentation of crude glycerol with mixed cultures in a simple medium

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Abstract

The objective of this study was to examine the applicability of mixed cultures for 1,3-propanediol (1,3-PDO) production from crude glycerol. Three different sources of mixed cultures were tested, where the mixed culture from a municipal wastewater treatment plant showed the best results. 1,3-PDO can be produced as the main product in this mixed culture with typical organic acids like acetic and butyric acids as by-products. The yield was in the range of 0.56–0.76 mol 1,3-PDO per mol glycerol consumed depending on the glycerol concentration. A final product concentration as high as 70 g/L was obtained in fed-batch cultivation with a productivity of 2.6 g/L h. 1,3-PDO can be kept in the culture several days after termination of the fermentation without being degraded. Degradation tests showed that 1,3-PDO is degraded much slower than other compounds in the fermentation broth. In comparison to 1,3-PDO production in typical pure cultures, the process developed in this work with a mixed culture achieved the same levels of product titer, yield and productivity, but has the decisive advantage of operation under complete non-sterile conditions. Moreover, a defined fermentation medium without yeast extract can be used and nitrogen gassing can be omitted during cultivation, leading to a strong reduction of investment and production costs.

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References

  1. Barbirato F, Camarasaclaret C, Grivet J, Bories A (1995) Glycerol fermentation by a new 1,3-propanediol producing microorganism—Enterobacter agglomerans. Appl Microbiol Biotechnol 43:786–793

    Article  CAS  Google Scholar 

  2. Barbirato F, Himmi E, Conte T, Bories A (1998) 1,3-Propanediol production by fermentation: an interesting way to valorize glycerin from the ester and ethanol industries. Ind Crops Prod 7:281–289

    Article  CAS  Google Scholar 

  3. Biebl H, Marten S, Hippe H, Deckwer W (1992) Glycerol conversion to 1,3-propanediol by newly isolated Clostridia. Appl Microbiol Biotechnol 36:592–597

    CAS  Google Scholar 

  4. Biebl H, Menzel K, Zeng A, Deckwer W (1999) Microbial production of 1,3-propanediol. Appl Microbiol Biotechnol 52:289–297

    Article  CAS  Google Scholar 

  5. Bizukojc M, Dietz D, Sun J, Zeng AP (2010) Metabolic modelling of syntrophic like growth of a 1,3-propanediol producer Clostridium butyricum and a methanogenic archeon Methanosarcina mazei under anaerobic conditions. Bioproc Biosyst Eng 33:507–552

    Article  CAS  Google Scholar 

  6. Chatzifragkou A, Papanikolaou S, Dietz D, Doulgeraki AI, Nychas GE, Zeng AP (2011) Production of 1,3-propanediol by Clostridium butyricum growing on biodiesel-derived crude glycerol through a non-sterilized fermentation process. Appl Microbiol Biotechnol 91:101–112

    Article  CAS  Google Scholar 

  7. Freund A (1881) Über die Bildung und Darstellung von Trimethylenalkohol aus Glycerin. Monatshefte Chemie 2:636–641

    Article  Google Scholar 

  8. Gonzalez-Pajuelo M, Meynial-Salles I, Mendes F, Soucaille P, Vasconcelos I (2006) Microbial conversion of glycerol to 1,3-propanediol: physiological comparison of a natural producer, Clostridium butyricum VPI 3266, and an engineered strain, Clostridium acetobutylicum DG1(pSPD5). Appl Microbiol Biotechnol 72:96–101

    CAS  Google Scholar 

  9. Himmi E, Bories A, Barbirato F (1999) Nutrient requirements for glycerol conversion to 1,3-propanediol by Clostridium butyricum. Bioresour Technol 67(2):123–128

    Article  CAS  Google Scholar 

  10. Hirschmann S, Baganz K, Koschik I, Vorlop KD (2005) Development of an integrated bioconversion process for the production of 1,3-propanediol from raw glycerol waters. Landbauforschung Völkenrode 55:261–267

    CAS  Google Scholar 

  11. Homann T, Tag C, Biebl H, Deckwer W, Schink B (1990) Fermentation of glycerol to 1,3- propanediol by Klebsiella and Citrobacter strains. Appl Microbiol Biotechnol 33:121–126

    CAS  Google Scholar 

  12. Majer S, Mueller-Langer F, Zeller V, Kaltschmitt M (2009) Implications of biodiesel production and utilisation on global climate—a literature review. Eur J Lipid Sci Technol 111:747–762

    Article  CAS  Google Scholar 

  13. Metsoviti M, Paraskeyajdi K, Koutinas A, Zeng AP, Papanikolaou S (2012) Production of 1,3-propanediol, 2,3-butanediol and ethanol by a newly isolated Klebsiella oxytoca strain growing on biodiesel-derived glycerol based media. Proc Biochem 47:1872–1882

    Article  CAS  Google Scholar 

  14. Metsoviti M, Zeng AP, Koutinas A, Papanikolaou S (2013) Enhanced 1,3-propanediiol production by a newly isolated Citrobacter freundii strain cultivated on biodiesel-derived waste glycerol through sterile and non-sterile bioprocesses. J Biotechnol 163:408–41815

    Article  CAS  Google Scholar 

  15. Oppenberg B, Schink B (1990) Anaerobic degradation of 1,3-propanediol by sulfate-reducing and by fermenting bacteria. Antonie van Leeuwenhoek 57:205–213

    Article  CAS  Google Scholar 

  16. Papanikolaou S, Ruiz-Sanchez P, Pariset B, Blanchard F, Fick M (2000) High production of 1,3-propanediol from industrial glycerol by a newly isolated Clostridium butyricum strain. J Biotechnol 77:191–208

    Article  CAS  Google Scholar 

  17. Papanikolaou S, Fick M, Aggelis G (2004) The effect of raw glycerol concentration on the production of 1,3-propanediol by Clostridium butyricum. J Chem Technol Biotechnol 79:1189–1196

    Article  CAS  Google Scholar 

  18. Pflugmacher U, Gottschalk G (1994) Development of an immobilized cell reactor for the production of 1,3-Propanediol by Citrobacter freundii. Appl Microbiol Biotechnol 41:313–316

    Article  CAS  Google Scholar 

  19. Qatibi A, Bories A, Garcia J (1991) Sulfate reduction and anaerobic glycerol degradation by a mixed microbial culture. Curr Microbiol 22:47–52

    Article  CAS  Google Scholar 

  20. Ringel AK, Wilkens E, Hortig D, Willke T, Vorlop KD (2012) An improved screening method for microorganisms able to convert crude glycerol to 1,3-propanediol and to tolerate high product concentrations. Appl Microbiol Biotechnol 93:1049–1056

    Article  CAS  Google Scholar 

  21. Sabra W, Dietz D, Tjahjasari D, Zeng AP (2010) Biosystems analysis and engineering of microbial consortia for industrial biotechnology. Eng Life Sci 10:407–421

    Article  CAS  Google Scholar 

  22. Saxena RK, Anand P, Saran S, Isar J (2009) Microbial production of 1,3-propanediol: recent developments and emerging opportunities. Biotechnol Adv 27:895–913

    Article  CAS  Google Scholar 

  23. Selembo PA, Perez JM, Lloyd WA, Logan BE (2009) Enhanced hydrogen and 1,3-propanediol production from glycerol by fermentation using mixed cultures. Biotechnol Bioeng 104:1098–1106

    Article  CAS  Google Scholar 

  24. Tanaka S, Lee Y (1997) Control of sulfate reduction by molybdate in anaerobic digestion. Water Sci Technol 36:143–150

    Article  CAS  Google Scholar 

  25. Temudo MF, Muyzer G, Kleerebezem R, van Loosdrecht MCM (2008) Diversity of microbial communities in open mixed culture fermentations: impact of the pH and carbon source. Appl Microbiol Biotechnol 80:1121–1130

    Article  CAS  Google Scholar 

  26. Temudo MF, Poldermans R, Kleerebezem R, van Loosdrecht MCM (2008) Glycerol fermentation by (open) mixed cultures: a chemostat study. Biotechnol Bioeng 100:1088–1098

    Article  CAS  Google Scholar 

  27. Tjahjasari D, Kaeding T, Zeng AP (2011) 1,3-Propanediol and Polytrimethyleneterephthalate. In: Moreina and Bai (eds) Comprehensive Biotechnology. vol 3: Ind Biotechnol Commodity Products, pp 229–242

  28. Wilkens E, Ringel AK, Hortig D, Willke T, Vorlop KD (2012) High-level production of 1,3-propanediol from crude glycerol by Clostridium butyricum AKR102a. Appl Microbiol Biotechnol 93:1057–1063

    Article  CAS  Google Scholar 

  29. Xiu ZL, Zeng AP (2008) Present state and perspective of downstream processing of biologically produced 1,3-propanediol and 2,3-butanediol. Appl Microbiol Biotechnol 78:917–926

    Article  CAS  Google Scholar 

  30. Zeng A (1995) A new balance equation for reducing equivalents for data consistency check and bioprocess calculation. J Biotechnol 43:111–124

    Article  CAS  Google Scholar 

  31. Zeng A (1996) Pathway and kinetic analysis of 1,3-propanediol production from glycerol fermentation by Clostridium butyricum. Bioprocess Eng 14:169–175

    Article  CAS  Google Scholar 

  32. Zeng A, Biebl H, Schlieker H, Deckwer W (1993) Pathway analysis of glycerol fermentation by Klebsiella pneumoniae - Regulation of reducing equivalent balance and product formation. Enyzme Microb Technol 15:770–779

    Article  CAS  Google Scholar 

  33. Zeng A, Ross A, Biebl H, Tag C, Gunzel B, Deckwer W (1994) Multiple product inhibition and growth modeling of Clostridium butyricum and Klebsiella pneumoniae in glycerol fermentation. Biotechnol Bioeng 44:902–911

    Article  CAS  Google Scholar 

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Acknowledgments

This work was financially supported by 7th Framework research program of the European Union through the project Propanergy (grant agreement no. 212671). We thank the project partners, especially Mr. Klaus Rohbrecht-Buck and Mr. Ralf Gabler from the company BKW Fürstenwalde for their help and support in the pilot plant fermentation.

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Correspondence to Donna Dietz.

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Dietz, D., Zeng, AP. Efficient production of 1,3-propanediol from fermentation of crude glycerol with mixed cultures in a simple medium. Bioprocess Biosyst Eng 37, 225–233 (2014). https://doi.org/10.1007/s00449-013-0989-0

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  • DOI: https://doi.org/10.1007/s00449-013-0989-0

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