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Impact of impurities in biodiesel-derived crude glycerol on the fermentation by Clostridium pasteurianum ATCC 6013

  • Bioenergy and biofuels
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Abstract

During the production of biodiesel, crude glycerol is produced as a byproduct at 10% (w/w). Clostridium pasteurianum has the inherent potential to grow on glycerol and produce 1,3-propanediol and butanol as the major products. Growth and product yields on crude glycerol were reported to be slower and lower, respectively, in comparison to the results obtained from pure glycerol. In this study, we analyzed the effect of each impurity present in the biodiesel-derived crude glycerol on the growth and metabolism of glycerol by C. pasteurianum. The crude glycerol contains methanol, salts (in the form of potassium chloride or sulfate), and fatty acids that were not transesterified. Salt and methanol were found to have no negative effects on the growth and metabolism of the bacteria on glycerol. The fatty acid with a higher degree of unsaturation, linoleic acid, was found to have strong inhibitory effect on the utilization of glycerol by the bacteria. The fatty acid with lower or no degrees of unsaturation such as stearic and oleic acid were found to be less detrimental to substrate utilization. The removal of fatty acids from crude glycerol by acid precipitation resulted in a fermentation behavior that is comparable to the one on pure glycerol. These results show that the fatty acids in the crude glycerol have a negative effect by directly affecting the utilization of glycerol as the carbon source, and hence their removal from crude glycerol is an essential step towards the utilization of crude glycerol.

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Reference

  • Atsumi S, Cann AF, Connor MR, Shen CR, Smith KM, Brynildsen MP, Chou KJY, Hanai T, Liao JC (2008) Metabolic engineering of Escherichia coli for 1-butanol production. Metab Eng 10:305–311

    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 53:2854–2861

    CAS  Google Scholar 

  • Baumann NA, Hagen PO, Goldfine H (1965) Phospholipids of Clostridium butyricum. J Biol Chem 240:1559–1567

    CAS  Google Scholar 

  • Biebl H (2001) Fermentation of glycerol by Clostridium pasteurianum—batch and continuous culture studies. J Ind Microbiol Biotechnol 27:18–26

    Article  CAS  Google Scholar 

  • Chi Z, Pyle D, Wen Z, Frear C, Chen S (2007) A laboratory study of producing docosahexaenoic acid from biodiesel-waste glycerol by microalgal fermentation. Process Biochem 42:1537–1545. doi:10.1016/j.procbio.2007.08.008

    Article  CAS  Google Scholar 

  • Dabrock B, Bahl H, Gottschalk G (1992) Parameters affecting solvent production by Clostridium pasteurianum. Appl Environ Microbiol 58:1233–1239

    CAS  Google Scholar 

  • Desbois AP, Smith VJ (2010) Antibacterial free fatty acids: activities, mechanisms of action and biotechnological potential. Appl Microbiol Biotechnol 85:1629–1642

    Article  CAS  Google Scholar 

  • Dharmadi Y, Murarka A, Gonzalez R (2006) Anaerobic fermentation of glycerol by Escherichia coli: a new platform for metabolic engineering. Biotechnol Bioeng 94:821–829

    Article  CAS  Google Scholar 

  • Furusawa H, Koyama N (2004) Effect of fatty acids on the membrane potential of an alkaliphilic bacillus. Curr Microbiol 48:196–198

    Article  CAS  Google Scholar 

  • Galbraith H, Miller TB (1973) Effect of long chain fatty acids on bacterial respiration and amino acid uptake. J Appl Microbiol 36:659–675

    Article  CAS  Google Scholar 

  • Gonzalez-Pajuelo M, Andrade JC, Vasconcelos I (2004) Production of 1, 3-propanediol by Clostridium butyricum VPI 3266 using a synthetic medium and raw glycerol. J Ind Microbiol Biotechnol 31:442–446

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Huang WC, Ramey DE, Yang ST (2004) Continuous production of butanol by Clostridium acetobutylicum immobilized in a fibrous bed bioreactor. Appl Biochem Biotechnol 115:887–898

    Article  Google Scholar 

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

    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 

  • Kabelitz N, Santos PM, Heipieper HJ (2003) Effect of aliphatic alcohols on growth and degree of saturation of membrane lipids in Acinetobacter calcoaceticus. FEMS Microbiol Lett 220:223–227

    Article  CAS  Google Scholar 

  • Khuller GK, Goldfine H (1975) Replacement of acyl an alk-1-enyl groups in Clostridium butyricum phospholipids by exogenous fatty acids. Biochem 16:3642–3647. doi:10.1021/bi00687a020

    Google Scholar 

  • Liang Y, Sarkany N, Cui Y, Blackburn JW (2010) Batch stage study of lipid production from crude glycerol derived from yellow grease or animal fats through microalgal fermentation. Bioresour Technol 101:6745–6750. doi:10.1016/j.biortech.2010.03.087

    Article  CAS  Google Scholar 

  • Muranushi N, Takagi N, Muranishi S, Sezaki H (1981) Effect of fatty acids and monoglycerides on permeability of lipid bilayer. Chem Phys Lipids 28:269–279

    Article  CAS  Google Scholar 

  • Pagliaro M, Ciriminna R, Kimura H, Rossi M, Della Pina C (2007) From glycerol to value added products. Angew Chem Int Ed Engl 46:4434–4440

    Article  CAS  Google Scholar 

  • Papanikolaou S, Muniglia L, Chevalot I, Aggelis G, Marc I (2002) Yarrowia lipolytica as a potential producer of citric acid from raw glycerol. J Appl Microbiol 92:737–744

    Article  CAS  Google Scholar 

  • Petrache HI, Tristram-Nagle S, Harries D, Kucerka N, Nagle JF, Parsegian VA (2006) Swelling of phospholipids by monovalent salt. J Lipid Res 47:302–309

    Article  CAS  Google Scholar 

  • Pyle DJ, Garcia RA, Wen Z (2008) Producing docosahexaenoic acid (DHA)-rich algae from biodiesel-derived crude glycerol: effects of impurities on DHA production and algal biomass composition. J Agr Food Chem 56:3933–3939

    Article  CAS  Google Scholar 

  • Raynaud C, Sarçabal P, Meynial-Salles I, Croux C, Soucaille P (2003) Molecular characterization of the 1, 3-propanediol (1, 3-PD) operon of Clostridium butyricum. Proc Natl Acad Sci USA 100:5010–5015

    Article  CAS  Google Scholar 

  • Taconi KA, Venkataramanan KP, Johnson DT (2009) Growth and solvent production by Clostridium pasteurianum ATCC® 6013TM utilizing biodiesel derived crude glycerol as the sole carbon source. Environ Prog Sustain Energy 28:100–110

    Article  CAS  Google Scholar 

  • Thompson L, Cockayne A, Spiller RC (1994) Inhibitory effect of polyunsaturated fatty acids on the growth of Helicobacter pylori: a possible explanation of the effect of diet on peptic ulceration. Gut 35:1557–1561

    Article  CAS  Google Scholar 

  • Urbanchuk JM (2011) Economic Impact of Removing the Biodiesel Tax Credit for 2010 and Implementation of RFS2 Targets Through 2015. Prepared for the National Biodiesel Board. June 8. Accessed 26 June 2011. http://www.biodiesel.org/resources/reportsdatabase/reports/gen/20110608_GEN-425.pdf.

  • Yazdani SS, Gonzalez R (2007) Anaerobic fermentation of glycerol: a path to economic viability for the biofuels industry. Curr Opin Biotechnol 18:213–219

    Article  CAS  Google Scholar 

  • Yokochi T, Honda D, Higashihara T, Nakahara T (1998) Optimization of docosahexaenoic acid production by Schizochytrium limacinum SR21. Appl Microbiol Biotechnol 49:72–76

    Article  CAS  Google Scholar 

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Acknowledgments

This project was supported by the NSF grant (CBET—0966846). We sincerely thank Tracy Armstrong for his help with NMR spectroscopy.

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Authors and Affiliations

  1. Department of Chemical and Materials Engineering, University of Alabama in Huntsville, 301 Sparkman Dr NW, MSB 333, Huntsville, AL, 35899, USA

    Keerthi P. Venkataramanan, Judy J. Boatman & Katherine A. Taconi

  2. Department of Chemical Engineering, University of Rhode Island, 203 Crawford Hall, Kingston, RI, 02881, USA

    Yogi Kurniawan & Geoffrey D. Bothun

  3. Department of Chemistry, University of Alabama in Huntsville, 301 Sparkman Dr NW, MSB 333, Huntsville, AL, 35899, USA

    Carmen Scholz

Authors
  1. Keerthi P. Venkataramanan
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  2. Judy J. Boatman
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  3. Yogi Kurniawan
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  4. Katherine A. Taconi
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  5. Geoffrey D. Bothun
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  6. Carmen Scholz
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Correspondence to Carmen Scholz.

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Venkataramanan, K.P., Boatman, J.J., Kurniawan, Y. et al. Impact of impurities in biodiesel-derived crude glycerol on the fermentation by Clostridium pasteurianum ATCC 6013. Appl Microbiol Biotechnol 93, 1325–1335 (2012). https://doi.org/10.1007/s00253-011-3766-5

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  • DOI: https://doi.org/10.1007/s00253-011-3766-5

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