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Adaptation dynamics of Clostridium butyricum in high 1,3-propanediol content media

  • Applied microbial and cell physiology
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Abstract

Aim of the present study was to evaluate the effect of exogenous additions of 1,3-propanediol (1,3-PDO) on microbial growth and metabolites production of Clostridium butyricum VPI 1718 strain, during crude glycerol fermentation. Preliminary batch cultures in anaerobic Duran bottles revealed that early addition of 1,3-PDO caused growth cessation in rather low quantities (15 g/L), while 1,3-PDO additions during the middle exponential growth phase up to 70 g/L resulted in an almost linear decrease of the specific growth rate (μ), accompanied by reduced glycerol assimilation, with substrate consumption being used mainly for energy of maintenance requirements. During batch trials in a 3-L bioreactor, the strain proved able to withstand more than 70 g/L of both biologically produced and externally added 1,3-PDO, whereas glycerol assimilation and metabolite production were carried on at a lower rate. Adaptation of the strain in high 1,3-PDO concentration environments was validated during its continuous cultivation with pulses of 1,3-PDO in concentrations of 31 and 46 g/L, where no washout phenomena were noticed. As far as C. butyricum cellular lipids were concerned, during batch bioreactor cultivations, 1,3-PDO addition was found to favor the biosynthesis of unsaturated fatty acids. Also, fatty acid composition was studied during continuous cultures, in which additions of 1,3-PDO were performed at steady states. Lipids were globally more saturated compared to batch cultures, while by monitoring of the transitory phases, it was noticed that the gradual diol washout had an evident impact in the alteration of the fatty acid composition, by rendering them more unsaturated.

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References

  • Abbad-Andaloussi S, Manginot-Dürr C, Amine J, Petitdemange E, Petitdemange H (1995) Isolation and characterization of Clostridium butyricum DSM 5431 mutants with increased resistance to 1,3-propanediol and altered production of acids. Appl Environ Microbiol 61:4413–4417

    CAS  Google Scholar 

  • AFNOR (1984) Recueil des normes françaises des corps gras, grains oléagineux et produis dérivés. In: Association Fraincaise pour Normalisation, Paris, p. 95

  • Amaral PF, Ferreira TF, Fontes GC, Coelho MAZ (2009) Glycerol valorization: New biotechnological routes. Food Bioprod Proc 87:179–186

    Article  CAS  Google Scholar 

  • Baer SH, Blaschek HP, Smith TL (1987) Effect of butanol challenge and temperature on lipid composition and membrane fluidity of butanol-tolerant Clostridium acetobutylicum. Appl Environ Microbiol 53(12):2854–2861

    CAS  Google Scholar 

  • Barbirato F, Himmi EH, Conte T, Bories A (1998) 1,3-propanediol production by fermentation: An interesting way to valorize glycerin from the ester and ethanol industries. Ind Crop Prod 7:281–289

    Article  CAS  Google Scholar 

  • Baumann NA, Hagen PO, Goldfine H (1964) Phospholipids of Clostridium butyricum. Studies on plasmalogen composition and biosynthesis. J Biol Chem 240(4):1559–1567

    Google Scholar 

  • Biebl H (1991) Glycerol fermentation of 1,3-propanediol by Clostridium butyricum. Measurement of product inhibition by use of a pH-auxostat. Appl Microbiol Biotechnol 35:701–705

    Article  CAS  Google Scholar 

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

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Biebl H, Spröer C (2002) Taxonomy of the glycerol fermenting Clostridia and description of Clostridium diolis sp. nov. Syst Appl Microbiol 25:491–497

    Article  CAS  Google Scholar 

  • Bloch K (1969) Enzymatic synthesis of monounsaturated fatty acids. Accounts Chem Res 2(7):193–202

    Article  CAS  Google Scholar 

  • Bloomfeld DK, Bloch K (1960) The formation of Δ9-unsaturated fatty acids. J Biol Chem 235:337–345

    Google Scholar 

  • Cameron DC, Altaras NE, Hoffman ML, Shaw AJ (1998) Metabolic engineering of propanediol pathways. Biotechnol Prog 14:116–125

    Article  CAS  Google Scholar 

  • Chatzifragkou A, Dietz D, Komaitis M, Zeng AP, Papanikolaou S (2010) Effect of biodiesel-derived waste glycerol impurities on biomass and 1,3-propanediol production of Clostridium butyricum VPI 1718. Biotechnol Bioeng 107:76–84

    Article  CAS  Google Scholar 

  • Chatzifragkou A, Aggelis G, Komaitis M, Zeng AP, Papanikolaou S (2011) Impact of anaerobiosis strategy and bioreactor geometry on the biochemical response of Clostridium butyricum VPI 1718 during 1,3-propanediol fermentation. Bioresour Technol 102:10625–10632

    Article  CAS  Google Scholar 

  • Colin T, Bories A, Moulin G (2000) Inhibition of Clostridium butyricum by 1,3-propanediol and diols during glycerol fermentation. Appl Microbiol Biotechnol 54:201–205

    Article  CAS  Google Scholar 

  • Deckwer WD (1995) Microbial conversion of glycerol to 1,3-propanediol. FEMS Microbiol Rev 16:143–149

    Article  CAS  Google Scholar 

  • Dobson G, Christie WW (2002) Mass spectrometry of fatty acid derivatives. Eur J Lipid Sci Technol 104:36–43

    Google Scholar 

  • Fulco AJ (1983) Fatty acid metabolism in bacteria. Prog Lipid Res 22:133–160

    Article  CAS  Google Scholar 

  • Goldfine H, Bloch K (1961) On the origin of unsaturated fatty acids in Clostridia. J Biol Chem 236(10):2596–2601

    CAS  Google Scholar 

  • 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. Landbauforsch Volk 55:261–267

    CAS  Google Scholar 

  • Ingram LO (1976) Adaptation of membrane lipids to alcohols. J Bacteriol 125:670–678

    CAS  Google Scholar 

  • Jain MK, Gleeson J, Upreti A, Upreti GC (1978) Intrinsic perturbing ability of akanols in lipid bilayers. Biochim Biophys Acta 509:1–8

    Article  CAS  Google Scholar 

  • Johnson DT, Taconi KA (2007) The glycerin glut: Options for the value-added conversion of crude glycerol resulting from biodiesel production. Environ Prog 26:338–348

    Article  CAS  Google Scholar 

  • Lepage C, Fayoulle F, Hermann M, Vandecasteele JP (1987) Changes in membrane lipid composition of Clostridium acetobutylicum during acetone-butanol fermentation: effects of solvents, growth temperature and pH. J Gen Microbiol 133:103–110

    CAS  Google Scholar 

  • Linden JC, Moreira A (1982) Anaerobic production of chemicals. In: Hollaender A (ed) Basic biology of new developments in biotechnology, vol 25, Basic life science. Plenum Press, New York, pp 377–403

    Google Scholar 

  • Menzel K, Zeng AP, Deckwer WD (1997) High concentration and productivity of 1,3-propanediol from continuous fermentation of glycerol by Klebsiella pneumoniae. Enzyme Microb Technol 20:82–86

    Article  CAS  Google Scholar 

  • Nakamura CE, Whited GM (2003) Metabolic engineering for the microbial production of 1,3-propanediol. Curr Opin Biotechnol 14:454–459

    Article  CAS  Google Scholar 

  • Nichols DS, Nichols PD, McMeekin TA (1992) Anaerobic production of polyunsaturated fatty acids by Shewanella putrefaciens strain ACAM 342. FEMS Microbiol Lett 98:117–122

    Article  CAS  Google Scholar 

  • 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 

  • 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 

  • Papanikolaou S, Fakas S, Fick M, Chevalot I, Galiotou-Panayotou M, Komaitis M, Marc I, Aggelis G (2008) Biotechnological valorization of raw glycerol discharged after bio-diesel (fatty acid methyl esters) manufacturing process: Production of 1,3-propanediol, citric acid and single cell oil. Biomass Bioenerg 32(1):60–71

    Article  CAS  Google Scholar 

  • Papanikolaou S (2009) Microbial conversion of glycerol into 1,3-propanediol: Glycerol assimilation, biochemical events related with 1,3-propanediol biosynthesis and biochemical engineering of the process. In: Aggelis G (ed) Microbial conversions of raw glycerol. Nova Science Publishers Inc., New York, pp 137–168

    Google Scholar 

  • Petitdemange E, Dürr C, Abbad Andaloussi S, Raval G (1995) Fermentation of raw glycerol to 1,3-propanediol by new strains of Clostridium butyricum. J Ind Microbiol 15:498–502

    Article  CAS  Google Scholar 

  • Reimann A, Biebl H (1996) Production of 1,3-propanediol by Clostridium butyricum DSM 5431 and product tolerant mutants in fed-batch culture: Feeding strategy for glycerol and ammonium. Biotechnol Lett 18:827–832

    Article  CAS  Google Scholar 

  • 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 

  • Scheuerbrandt G, Bloch K (1962) Unsaturated fatty acids in microorganisms. J Biol Chem 237(7):2064–2068

    CAS  Google Scholar 

  • Shimizu T, Katsura T (1988) Steady-state kinetic study on the inhibition of the adenosinetriphosphatase activity of dynein from Tetrahymena cilia by glycerol. J Biochem 103:99–105

    CAS  Google Scholar 

  • Vetter W, Thurnhofer S (2007) Analysis of fatty acids by mass spectrometry in the selected ion monitoring mode. Lipid Technol 19:184–186

    Article  CAS  Google Scholar 

  • Vollherbst-Schneck K, Sands JA, Montenecourt BS (1984) Effect of butanol on lipid composition and fluidity of Clostridium acetobutylicum ATCC 824. Appl Environ Microbiol 47:193–194

    CAS  Google Scholar 

  • 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 

  • Willke T, Vorlop KD (2008) Biotransformation of glycerol into 1,3-propanediol. Eur J Lipid Sci Technol 110:831–840

    Article  CAS  Google Scholar 

  • Zeng AP, Ross A, Biebl H, Tag C, Günzel B, Deckwer WD (1994) Multiple product inhibition and growth modeling of Clostridium butyricum and Klebsiella pneumoniae in glycerol fermentation. Biotechnol Bioeng 44:902–922

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Zeng AP, Biebl H (2002) Bulk chemicals from biotechnology: The case of 1,3-propanediol production and the new trends. Adv Biochem Eng Biotechnol 74:239–259

    CAS  Google Scholar 

  • Zeng AP, Sabra W (2011) Microbial production of diols as platform chemicals: Recent progresses. Curr Opin Biotechnol 22:749–757

    Article  CAS  Google Scholar 

  • Zhang YM, Rock CO (2008) Membrane lipid homeostasis in bacteria. Nat Rev Microbiol 6:222–233

    Article  Google Scholar 

Download references

Acknowledgments

Financial support has been provided by the EU (FP7 Program “PROPANERGY—Integrated bioconversion of glycerine into value-added products and biogas at pilot plant scale”, Grant number: 212671).

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Authors and Affiliations

  1. Department of Food Science and Technology, Agricultural University of Athens, 75 Iera Odos, 11855, Athens, Greece

    Afroditi Chatzifragkou, Chryssavgi Gardeli, Maria Galiotou-Panayotou, Michael Komaitis & Seraphim Papanikolaou

  2. Unit of Microbiology, Department of Biology, Division of Genetics, Cell and Development Biology, University of Patras, 26504, Patras, Greece

    George Aggelis

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  1. Afroditi Chatzifragkou
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  2. George Aggelis
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  3. Chryssavgi Gardeli
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  4. Maria Galiotou-Panayotou
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  6. Seraphim Papanikolaou
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Correspondence to Seraphim Papanikolaou.

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This manuscript is dedicated to the memory of our beloved friend and colleague Professor Maria Galiotou, who passed away on January 21, 2012.

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Chatzifragkou, A., Aggelis, G., Gardeli, C. et al. Adaptation dynamics of Clostridium butyricum in high 1,3-propanediol content media. Appl Microbiol Biotechnol 95, 1541–1552 (2012). https://doi.org/10.1007/s00253-012-4003-6

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  • DOI: https://doi.org/10.1007/s00253-012-4003-6

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