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.
Similar content being viewed by others
References
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
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
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
Biebl H, Menzel K, Zeng A, Deckwer W (1999) Microbial production of 1,3-propanediol. Appl Microbiol Biotechnol 52:289–297
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
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
Freund A (1881) Über die Bildung und Darstellung von Trimethylenalkohol aus Glycerin. Monatshefte Chemie 2:636–641
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
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
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
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
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
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
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
Oppenberg B, Schink B (1990) Anaerobic degradation of 1,3-propanediol by sulfate-reducing and by fermenting bacteria. Antonie van Leeuwenhoek 57:205–213
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
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
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
Qatibi A, Bories A, Garcia J (1991) Sulfate reduction and anaerobic glycerol degradation by a mixed microbial culture. Curr Microbiol 22:47–52
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
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
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
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
Tanaka S, Lee Y (1997) Control of sulfate reduction by molybdate in anaerobic digestion. Water Sci Technol 36:143–150
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
Temudo MF, Poldermans R, Kleerebezem R, van Loosdrecht MCM (2008) Glycerol fermentation by (open) mixed cultures: a chemostat study. Biotechnol Bioeng 100:1088–1098
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
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
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
Zeng A (1995) A new balance equation for reducing equivalents for data consistency check and bioprocess calculation. J Biotechnol 43:111–124
Zeng A (1996) Pathway and kinetic analysis of 1,3-propanediol production from glycerol fermentation by Clostridium butyricum. Bioprocess Eng 14:169–175
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
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
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-013-0989-0