Stability and oscillatory behavior of microbial consortium in continuous conversion of crude glycerol to 1,3-propanediol

Abstract

Microbial consortium is an alternative for bioconversion of crude glycerol to value-added products whereas concerns about the process stability in long-term operation existed. The aim of this study is to evaluate the feasibility of using an anaerobic microbial consortium as inoculum for continuous conversion of crude glycerol to 1,3-propanediol (1,3-PDO). Performances of continuous fermentations with the consortium inoculum were evaluated under different dilution rates and glycerol feed concentrations. The highest 1,3-PDO production of 57.86 g/L was achieved with a productivity of 5.55 g/(L·h). Analyses of kinetic data showed that the consortium maintained a consistent pattern for 1,3-PDO production under different operating conditions despite changes in community composition. The continuous fermentation by the consortium was able to operate for a longer period of time (31 volume changes) than that using pure culture (24 volume changes) with the average 1,3-PDO concentration of 53.52 g/L and productivity of 6.69 g/(L·h) under glycerol-excess condition, which could be contributed to the intraspecies diversity among Clostridium butyricum in the consortium. Under glycerol-limited conditions, however, a spontaneous oscillation of the consortium was observed after continuous operation for about 120 h, along with severe fluctuations of the microbial community. The oscillatory behavior could be reduced by increasing the dilution rates and was likely the metabolic feature of C. butyricum.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

References

  1. Abbad-Andaloussi S, Guedon E, Spiesser E, Petitdemange H (1996) Glycerol dehydratase activity: the limiting step for 1,3-propanediol production by Clostridium butyricum DSM 5431. Lett Appl Microbiol 22(4):311–314. https://doi.org/10.1111/j.1472-765X.1996.tb01168.x

    CAS  Article  Google Scholar 

  2. Agler MT, Wrenn BA, Zinder SH, Angenent LT (2011) Waste to bioproduct conversion with undefined mixed cultures: the carboxylate platform. Trends Biotechnol 29(2):70–78. https://doi.org/10.1016/j.tibtech.2010.11.006

    CAS  Article  PubMed  Google Scholar 

  3. Almeida JR, Fávaro LC, Quirino BF (2012) Biodiesel biorefinery: opportunities and challenges for microbial production of fuels and chemicals from glycerol waste. Biotechnol Biofuels 5(1):48. https://doi.org/10.1186/1754-6834-5-48

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. Balagadde FK, Song H, Ozaki J, Collins CH, Barnet M, Arnold FH, Quake SR, You L (2008) A synthetic Escherichia coli predator-prey ecosystem. Mol Syst Biol 4(1):187. https://doi.org/10.1038/msb.2008.24

    Article  PubMed  PubMed Central  Google Scholar 

  5. 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(6):701–705. https://doi.org/10.1007/BF00169880

    CAS  Article  Google Scholar 

  6. Chatzifragkou A, Papanikolaou S, Dietz D, Doulgeraki AI, Nychas GJ, 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(1):101–112. https://doi.org/10.1007/s00253-011-3247-x

    CAS  Article  PubMed  Google Scholar 

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

    CAS  Article  PubMed  Google Scholar 

  8. Coyte KZ, Schluter J, Foster KR (2015) The ecology of the microbiome: networks, competition, and stability. Science 350(6261):663–666. https://doi.org/10.1126/science.aad2602

    CAS  Article  PubMed  Google Scholar 

  9. Dietz D, Zeng AP (2014) Efficient production of 1,3-propanediol from fermentation of crude glycerol with mixed cultures in a simple medium. Bioprocess Biosyst Eng 37(2):225–233. https://doi.org/10.1007/s00449-013-0989-0

    CAS  Article  PubMed  Google Scholar 

  10. Gallardo R, Faria C, Rodrigues LR, Pereira MA, Alves MM (2014) Anaerobic granular sludge as a biocatalyst for 1,3-propanediol production from glycerol in continuous bioreactors. Bioresour Technol 155:28–33. https://doi.org/10.1016/j.biortech.2013.12.008

    CAS  Article  PubMed  Google Scholar 

  11. Gonzalez-Pajuelo M, Andrade JC, Vasconcelos I (2005) Production of 1,3-propanediol by Clostridium butyricum VPI 3266 in continuous cultures with high yield and productivity. J Ind Microbiol Biotechnol 32(9):391–396. https://doi.org/10.1007/s10295-005-0012-0

    CAS  Article  PubMed  Google Scholar 

  12. 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 Environ Microbiol 72(1):96–101. https://doi.org/10.1128/AEM.72.1.96-101.2006

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. Hoskisson PA, Hobbs G (2005) Continuous culture—making a comeback? Microbiology 151(10):3153–3159. https://doi.org/10.1099/mic.0.27924-0

    CAS  Article  PubMed  Google Scholar 

  14. Jiang LL, Zhou JJ, Xiu ZL (2017a) Advances in industrial microbiome based on microbial consortium for biorefinery. Bioresour Bioprocess 4(1):11. https://doi.org/10.1186/s40643-017-0141-0

    Article  PubMed  PubMed Central  Google Scholar 

  15. Jiang LL, Liu HF, Mu Y, Sun YQ, Xiu ZL (2017b) High tolerance to glycerol and high production of 1,3-propanediol in batch fermentations by microbial consortium from marine sludge. Eng Life Sci 17(6):635–644. https://doi.org/10.1002/elsc.201600215

    CAS  Article  Google Scholar 

  16. Jiang LL, Dai JY, Sun YQ, Xiu ZL (2018) The effects of ionic liquid 1-ethyl-3-methylimidazolium trifluoromethanesulfonate on 1,3-propanediol production from crud glycerol by microbial consortium. Bioprocess Biosyst Eng. https://doi.org/10.1007/s00449-018-1937-9

    CAS  Article  PubMed  Google Scholar 

  17. Kanjilal B, Noshadi I, Bautista EJ, Srivastava R, Parnas RS (2015) Batch, design optimization, and DNA sequencing study for continuous 1,3-propanediol production from waste glycerol by a soil-based inoculum. Appl Microbiol Biotechnol 99(5):2105–2117. https://doi.org/10.1007/s00253-014-6259-5

    CAS  Article  PubMed  Google Scholar 

  18. Kaur G, Srivastava AK, Chand S (2012) Advances in biotechnological production of 1,3-propanediol. Biochem Eng J 64:106–118. https://doi.org/10.1016/j.bej.2012.03.002

    CAS  Article  Google Scholar 

  19. Kleerebezem R, van Loosdrecht MC (2007) Mixed culture biotechnology for bioenergy production. Curr Opin Biotechnol 18(3):207–212. https://doi.org/10.1016/j.copbio.2007.05.001

    CAS  Article  PubMed  Google Scholar 

  20. Lee CS, Aroua MK, Daud WMAW, Cognet P, Pérès-Lucchese Y, Fabre PL, Reynes O, Latapie L (2015) A review: conversion of bioglycerol into 1,3-propanediol via biological and chemical method. Renew Sust Energ Rev 42:963–972. https://doi.org/10.1016/j.rser.2014.10.033

    CAS  Article  Google Scholar 

  21. Li T, Chen XB, Chen JC, Wu Q, Chen GQ (2014) Open and continuous fermentation: products, conditions and bioprocess economy. Biotechnol J 9(12):1503–1511. https://doi.org/10.1002/biot.201400084

    CAS  Article  PubMed  Google Scholar 

  22. Liu H, Xu Y, Zheng Z, Liu D (2010) 1,3-Propanediol and its copolymers: research, development and industrialization. Biotechnol J 5(11):1137–1148. https://doi.org/10.1002/biot.201000140

    CAS  Article  PubMed  Google Scholar 

  23. Loureiro-Pinto M, Coca M, González-Benito G, Lucas S, García-Cubero MT (2017) Continuous bioproduction of 1,3-propanediol from biodiesel raw glycerol: operation with free and immobilized cells of Clostridium butyricum DSM 10702. Can J Chem Eng 95(5):819–826. https://doi.org/10.1002/cjce.22725

    CAS  Article  Google Scholar 

  24. Luedeking R, Piret EL (1959) A kinetic study of the lactic acid fermentation. Batch process at controlled pH. Biotechnol Bioeng 1(4):393–412. https://doi.org/10.1002/jbmte.390010406

    CAS  Article  Google Scholar 

  25. Menzel K, Zeng AP, Biebl H, Deckwer WD (1996) Kinetic, dynamic, and pathway studies of glycerol metabolism by Klebsiella pneumoniae in anaerobic continuous culture: I. The phenomena and characterization of oscillation and hysteresis. Biotechnol Bioeng 52(5):549–560. https://doi.org/10.1002/(SICI)1097-0290(19961205)52:5<549::AID-BIT2>3.0.CO;2-J

    CAS  Article  PubMed  Google Scholar 

  26. Menzel K, Zeng AP, Deckwer WD (1997) High concentration and productivity of 1,3-propanediol from continuous fermentation of glycerol by Klebsiella pneumoniae. Enzym Microb Technol 20(2):82–86. https://doi.org/10.1016/S0141-0229(96)00087-7

    CAS  Article  Google Scholar 

  27. Monteiro MR, Kugelmeier CL, Pinheiro RS, Batalha MO, da Silva CA (2018) Glycerol from biodiesel production: technological paths for sustainability. Renew Sust Energ Rev 88:109–122. https://doi.org/10.1016/j.rser.2018.02.019

    CAS  Article  Google Scholar 

  28. Mu Y, Teng H, Zhang DJ, Wang W, Xiu ZL (2006) Microbial production of 1,3-propanediol by Klebsiella pneumoniae using crude glycerol from biodiesel preparations. Biotechnol Lett 28:1755–1759. https://doi.org/10.1007/s10529-006-9154-z

    CAS  Article  PubMed  Google Scholar 

  29. Mu XJ, Teng H, Sun YQ, Xiu ZL (2009) Techno-economic analysis on bioconversion of glycerol into 1,3-propanediol. Chin J Process Eng 9(5):947–952

    CAS  Google Scholar 

  30. 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(2):191–208. https://doi.org/10.1016/S0168-1656(99)00217-5

    CAS  Article  PubMed  Google Scholar 

  31. 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(11):1189–1196. https://doi.org/10.1002/jctb.1103

    CAS  Article  Google Scholar 

  32. Papanikolaou S, Rontou M, Belka A, Athenaki M, Gardeli C, Mallouchos A, Kalantzi O, Koutinas A, Kookos I, Zeng AP, Aggelis G (2017) Conversion of biodiesel-derived glycerol into biotechnological products of industrial significance by yeast and fungal strains. Eng Life Sci 17(3):262–281. https://doi.org/10.1002/elsc.201500191

    CAS  Article  Google Scholar 

  33. Petitdemange E, Dürr C, Andaloussi SA, Raval G (1995) Fermentation of raw glycerol to 1,3-propanediol by new strains of Clostridium butyricum. J Ind Microbiol 15(6):498–502. https://doi.org/10.1007/BF01570021

    CAS  Article  Google Scholar 

  34. Pirt SJ (1965) The maintenance energy of bacteria in growing cultures. Proc R Soc Lond B 163(991):224–231. https://doi.org/10.1098/rspb.1965.0069

    CAS  Article  PubMed  Google Scholar 

  35. Reimann A, Abbad-Andaloussi S, Biebl H, Petitdemange H (1998) 1,3-Propanediol formation with product-tolerant mutants of Clostridium butyricum DSM 5431 in continuous culture: productivity, carbon and electron flow. J Appl Microbiol 84(6):1125–1130. https://doi.org/10.1046/j.1365-2672.1998.00453.x

    CAS  Article  Google Scholar 

  36. Ringel AK, Wilkens E, Hortig D, Willke T, Vorlop KD (2012) An improved screening method for microorganisms able to convert crudeglycerol to 1,3-propanediol and to tolerate high product concentrations. Appl Microbiol Biotechnol 93:1049–1056. https://doi.org/10.1007/s00253-011-3594-7

    CAS  Article  PubMed  Google Scholar 

  37. Rywińska A, Juszczyk P, Wojtatowicz M, Robak M, Lazar Z, Tomaszewska L, Rymowicz W (2013) Glycerol as a promising substrate for Yarrowia lipolytica biotechnological applications. Biomass Bioenergy 48:148–166. https://doi.org/10.1016/j.biombioe.2012.11.021

    CAS  Article  Google Scholar 

  38. Sabra W, Dietz D, Tjahjasari D, Zeng AP (2010) Biosystems analysis and engineering of microbial consortia for industrial biotechnology. Eng Life Sci 10(5):407–421. https://doi.org/10.1002/elsc.201000111

    CAS  Article  Google Scholar 

  39. Samul D, Leja K, Grajek W (2014) Impurities of crude glycerol and their effect on metabolite production. Ann Microbiol 64:891–898. https://doi.org/10.1007/s13213-013-0767-x

    CAS  Article  PubMed  Google Scholar 

  40. Sittijunda S, Reungsang A (2017) Fermentation of hydrogen, 1,3-propanediol and ethanol from glycerol as affected by organic loading rate using up-flow anaerobic sludge blanket (UASB) reactor. Int J Hydrog Energy 42(45):27558–27569. https://doi.org/10.1016/j.ijhydene.2017.05.149

    CAS  Article  Google Scholar 

  41. Sun LH, Song ZY, Sun YQ, Xiu ZL (2010) Dynamic behavior of glycerol–glucose co-fermentation for 1,3-propanediol production by Klebsiella pneumoniae DSM 2026 under micro-aerobic conditions. World J Microbiol Biotechnol 26(8):1401–1407. https://doi.org/10.1007/s11274-010-0314-x

    CAS  Article  Google Scholar 

  42. Sun YQ, Ma CW, Fu HX, Mu Y, Xiu ZL (2014) 1,3-Propanediol. In: Bisaria VS, Kondo A (eds) Bioprocessing of renewable resources to commodity bioproducts, 1st edn. John Wiley & Sons, Hoboken, pp 289–326

    Google Scholar 

  43. Sun YQ, Shen JT, Yan L, Zhou JJ, Jiang LL, Chen Y, Yuan JL, Feng EM, Xiu ZL (2018) Advances in bioconversion of glycerol to 1,3-propanediol: prospects and challenges. Process Biochem. https://doi.org/10.1016/j.procbio.2018.05.009

    CAS  Article  Google Scholar 

  44. Tomaszewska L, Rywińska A, Gładkowski W (2012) Production of erythritol and mannitol by Yarrowia lipolytica yeast in media containing glycerol. J Ind Microbiol Biotechnol 39(9):1333–1343. https://doi.org/10.1007/s10295-012-1145-6

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  45. Van Andel JG, Zoutberg GR, Crabbendam PM, Breure AM (1985) Glucose fermentation by Clostridium butyricum grown under a self generated gas atmosphere in chemostat culture. Appl Microbiol Biotechnol 23(1):21–26. https://doi.org/10.1007/BF02660113

    Article  Google Scholar 

  46. Varrone C, Floriotis G, Heggeset TMB, Le SB, Markussen S, Skiadas IV, Gavala HN (2017) Continuous fermentation and kinetic experiments for the conversion of crude glycerol derived from second-generation biodiesel into 1,3-propanediol and butyric acid. Biochem Eng J 128:149–161. https://doi.org/10.1016/j.bej.2017.09.012

    CAS  Article  Google Scholar 

  47. Wang X, Zhang H, Yang F, Wang Y, Gao M (2008) Long-term storage and subsequent reactivation of aerobic granules. Bioresour Technol 99(17):8304–8309. https://doi.org/10.1016/j.biortech.2008.03.024

    CAS  Article  PubMed  Google Scholar 

  48. Willke T, Vorlop K (2008) Biotransformation of glycerol into 1,3-propanediol. Eur J Lipid Sci Tech 110(9):831–840. https://doi.org/10.1002/ejlt.200800057

    CAS  Article  Google Scholar 

  49. Xiu ZL, Zeng AP (1999) A study progress in oscillatory and chaotic behavior in microbial continuous cultures. Prog Biotechnol 19(6):58–63

    Google Scholar 

  50. 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(6):917–926. https://doi.org/10.1007/s00253-008-1387-4

    CAS  Article  PubMed  Google Scholar 

  51. Zeng AP (1996) Pathway and kinetic analysis of 1,3-propanediol production from glycerol fermentation by Clostridium butyricum. Bioprocess Eng 14(4):169–175. https://doi.org/10.1007/BF01464731

    CAS  Article  Google Scholar 

  52. Zeng AP, Biebl H (2002) Bulk chemicals from biotechnology: the case of 1,3-propanediol production and the new trends. In: Tools and applications of biochemical engineering science. Springer, Berlin, pp 239–259

    Google Scholar 

  53. 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 gIyceroI fermentation. Biotechnol Bioeng 44(8):902–911. https://doi.org/10.1002/bit.260440806

    CAS  Article  PubMed  Google Scholar 

  54. Zeng AP, Menzel K, Deckwer W-D (1996) Kinetic, dynamic, and pathway studies of glycerol metabolism by Klebsiella pneumoniae in anaerobic continuous culture: II. Analysis of metabolic rates and pathways under oscillation and steady-state conditions. Biotechnol Bioeng 52(5):561–571. https://doi.org/10.1002/(SICI)1097-0290(19961205)52:5<561::AID-BIT3>3.0.CO;2-H

    CAS  Article  PubMed  Google Scholar 

  55. Zhang C, Yang H, Yang F, Ma Y (2009) Current progress on butyric acid production by fermentation. Curr Microbiol 59(6):656–663. https://doi.org/10.1007/s00284-009-9491-y

    CAS  Article  PubMed  Google Scholar 

  56. Zhou JJ, Shen JT, Jiang LL, Sun YQ, Mu Y, Xiu ZL (2017) Selection and characterization of an anaerobic microbial consortium with high adaptation to crude glycerol for 1,3-propanediol production. Appl Microbiol Biotechnol 101(15):5985–5996. https://doi.org/10.1007/s00253-017-8311-8

    CAS  Article  PubMed  Google Scholar 

  57. Zhu C, Chen B, Fang B (2013) Pretreatment of raw glycerol with activated carbon for 1,3-propanediol production by Clostridium butyricum. Eng Life Sci 13(4):376–384. https://doi.org/10.1002/elsc.201200106

    CAS  Article  Google Scholar 

Download references

Funding

This work was supported by the National Natural Science Foundation of China (Grant No. 21476042) and the Fundamental Research Funds for the Central Universities (Grant No. DUT17ZD209).

Author information

Affiliations

Authors

Corresponding author

Correspondence to Zhi-Long Xiu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Electronic supplementary material

ESM 1

(PDF 81 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhou, JJ., Shen, JT., Wang, XL. et al. Stability and oscillatory behavior of microbial consortium in continuous conversion of crude glycerol to 1,3-propanediol. Appl Microbiol Biotechnol 102, 8291–8305 (2018). https://doi.org/10.1007/s00253-018-9244-6

Download citation

Keywords

  • 1,3-Propanediol
  • Microbial consortium
  • Continuous fermentation
  • Clostridium butyricum
  • Stability
  • Oscillation