Skip to main content

Advertisement

SpringerLink
Log in

Physicochemical pretreatment selects microbial communities to produce alcohols through metabolism of volatile fatty acids

  • Original Article
  • Published:
Biomass Conversion and Biorefinery Aims and scope Submit manuscript

Abstract

This work reports the effect of four different physicochemical pretreatments (acidic, thermal, acidic-thermal, and thermal-acidic) on an anaerobic inoculum aiming at alcohols production, using acetate and butyrate as carbon sources and hydrogen as co-substrate. Pretreatments were carried out to select microbial communities more able to use hydrogen to metabolize volatile fatty acids into their respective alcohols. Experiments were conducted in single batches using acetate and butyrate as substrates at 30 °C and with a pressurized headspace of pure H2 at 2.15 atm (218.2 MPa). Thermal and acidic-thermal pretreatments lead to higher production of both ethanol and butanol, indicating that these pretreatments successfully selected communities more suitable for acetate and butyrate solventogenesis. Kinetics modelling shows that the highest attainable concentrations of ethanol and butanol produced were 122 mg L−1 and 97 mg L−1 for the thermal pretreatment (after 17.5 days) and 87 mg L−1 and 143 mg L−1 for the acidic-thermal pretreatment (after 18.9 days). Process thermodynamics indicated that high H2 partial pressure favoured solventogenic metabolic pathways. Sequencing data showed that both thermal and acidic-thermal pretreatments selected mainly the bacterial genera Pseudomonas, Brevundimonas, and Clostridium. Acidic-thermal pretreatment selected a bacterial community more adapted to the conversion of acetate and butyrate into ethanol and butanol, respectively. Thermal-acidic pretreatment was unstable, showing significant variability between replicates. Acidic pretreatment showed the lowest alcohol production.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Data availability

The datasets generated during and/or analysed during the current study are openly and freely available. All data, results, and data treatment, in the format of spreadsheets and downloadable files, used are published at Mendeley Data (https://doi.org/10.17632/wwm4zwbgrc.1). All sequences (and their processing) related to the 16S rRNA used to identify the microbial communities are available at MG-RAST (www.mg-rast.org/linkin.cgi?project=mgp10094).

References

  1. Jones DT, Woods DR (1986) Acetone-butanol fermentation revisited. Microbiol Rev 50:484–524

    Article  Google Scholar 

  2. Das D, Veziroglu T (2008) Advances in biological hydrogen production processes. Int J Hydrogen Energy 33:6046–6057. https://doi.org/10.1016/j.ijhydene.2008.07.098

    Article  Google Scholar 

  3. Baumann I, Westermann P (2016) Microbial production of short chain fatty acids from lignocellulosic biomass: current processes and market. BioMed Research International 2016

  4. Levin D (2004) Biohydrogen production: prospects and limitations to practical application. Int J Hydrogen Energy 29:173–185. https://doi.org/10.1016/S0360-3199(03)00094-6

    Article  Google Scholar 

  5. Steinbusch KJJ, Hamelers HVM, Buisman CJN (2008) Alcohol production through volatile fatty acids reduction with hydrogen as electron donor by mixed cultures. Water Res 42:4059–4066. https://doi.org/10.1016/j.watres.2008.05.032

    Article  Google Scholar 

  6. Zverlov VV, Berezina O, Velikodvorskaya G, a Schwarz WH (2006) Bacterial acetone and butanol production by industrial fermentation in the Soviet Union: use of hydrolyzed agricultural waste for biorefinery. Appl Microbiol Biotechnol 71:587–597. https://doi.org/10.1007/s00253-006-0445-z

    Article  Google Scholar 

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

    Article  Google Scholar 

  8. Dürre P (1998) New insights and novel developments in clostridial acetone/butanol/isopropanol fermentation. Applied Microbiology and Biotechnology 639–648

  9. Brynjarsdottir H, Wawiernia B, Orlygsson J (2012) Ethanol production from sugars and complex biomass by thermoanaerobacter AK 5: the effect of electron-scavenging systems on end-product formation. Energy Fuels 26:4568–4574

    Article  Google Scholar 

  10. Jessen JE, Orlygsson J (2012) Production of ethanol from sugars and lignocellulosic biomass by Thermoanaerobacter J1 isolated from a hot spring in Iceland. J Biomed Biotechnol 2012:186982. https://doi.org/10.1155/2012/186982

    Article  Google Scholar 

  11. Almarsdottir AR, Sigurbjornsdottir MA, Orlygsson J (2012) Effect of various factors on ethanol yields from lignocellulosic biomass by Thermoanaerobacterium AK17. Biotechnol Bioeng 109:686–694. https://doi.org/10.1002/bit.24346

    Article  Google Scholar 

  12. Crespo CF, Badshah M, Alvarez MT, Mattiasson B (2012) Ethanol production by continuous fermentation of D-(+)-cellobiose, D-(+)-xylose and sugarcane bagasse hydrolysate using the thermoanaerobe Caloramator boliviensis. Biores Technol 103:186–191. https://doi.org/10.1016/j.biortech.2011.10.020

    Article  Google Scholar 

  13. Xu L, Tschirner U (2011) Improved ethanol production from various carbohydrates through anaerobic thermophilic co-culture. Biores Technol 102:10065–10071. https://doi.org/10.1016/j.biortech.2011.08.067

    Article  Google Scholar 

  14. Mes-Hartree M, Saddler J (1982) Butanol production of Clostridium acetobutylicum grown on sugars found in hemicellulose hydrolysates. Biotech Lett 4:247–252

    Article  Google Scholar 

  15. Maddox I (1982) Production of ethanol and n-butanol from hexose/pentose mixtures using consecutive fermentations with Saccharomyces cerevisiae and Clostridium acetobutylicum. Biotech Lett 4:23–28

    Article  Google Scholar 

  16. Ounine K, Petitdemange H, Raval G, Gay R (1983) Acetone-butanol production from pentoses by Clostridium acetobutylicum. Biotech Lett 5:605–610

    Article  Google Scholar 

  17. Li Z, Shi Z, Li X (2014) Models construction for acetone-butanol-ethanol fermentations with acetate/butyrate consecutively feeding by graph theory. Biores Technol 159:320–326. https://doi.org/10.1016/j.biortech.2014.02.095

    Article  Google Scholar 

  18. Kumar M, Goyal Y, Sarkar A, Gayen K (2012) Comparative economic assessment of ABE fermentation based on cellulosic and non-cellulosic feedstocks. Appl Energy 93:193–204. https://doi.org/10.1016/j.apenergy.2011.12.079

    Article  Google Scholar 

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

    Article  Google Scholar 

  20. Puig S, Coma M, Monclús H et al (2008) Selection between alcohols and volatile fatty acids as external carbon sources for EBPR. Water Res 42:557–566. https://doi.org/10.1016/j.watres.2007.07.050

    Article  Google Scholar 

  21. O-Thong S, Prasertsan P, Birkeland N-K, (2009) Evaluation of methods for preparing hydrogen-producing seed inocula under thermophilic condition by process performance and microbial community analysis. Biores Technol 100:909–918. https://doi.org/10.1016/j.biortech.2008.07.036

    Article  Google Scholar 

  22. Luo G, Karakashev D, Xie L et al (2011) Long-term effect of inoculum pretreatment on fermentative hydrogen production by repeated batch cultivations: homoacetogenesis and methanogenesis as competitors to hydrogen production. Biotechnol Bioeng 108:1816–1827. https://doi.org/10.1002/bit.23122

    Article  Google Scholar 

  23. Pendyala B, Chaganti SR, Lalman J, a, et al (2012) Pretreating mixed anaerobic communities from different sources: correlating the hydrogen yield with hydrogenase activity and microbial diversity. Int J Hydrogen Energy 37:12175–12186. https://doi.org/10.1016/j.ijhydene.2012.05.105

    Article  Google Scholar 

  24. Kumar M, Gayen K, Saini S (2013) Role of extracellular cues to trigger the metabolic phase shifting from acidogenesis to solventogenesis in Clostridium acetobutylicum. Biores Technol 138:55–62. https://doi.org/10.1016/j.biortech.2013.03.159

    Article  Google Scholar 

  25. Angelidaki I, Petersen SP, Ahring BK (1990) Applied Microbiolog . v Effects of lipids on thermophilic anaerobic digestion and reduction of lipid inhibition upon addition of bentonite. 469–472

  26. Luo G, Xie L, Zou Z et al (2010) Evaluation of pretreatment methods on mixed inoculum for both batch and continuous thermophilic biohydrogen production from cassava stillage. Biores Technol 101:959–964. https://doi.org/10.1016/j.biortech.2009.08.090

    Article  Google Scholar 

  27. Mohan SV, Babu VL, Sarma PN (2008) Effect of various pretreatment methods on anaerobic mixed microflora to enhance biohydrogen production utilizing dairy wastewater as substrate 99:59–67. https://doi.org/10.1016/j.biortech.2006.12.004

    Article  Google Scholar 

  28. Dessì P, Porca E, Frunzo L et al (2018) Inoculum pretreatment differentially affects the active microbial community performing mesophilic and thermophilic dark fermentation of xylose. Int J Hydrogen Energy 43:9233–9245. https://doi.org/10.1016/j.ijhydene.2018.03.117

    Article  Google Scholar 

  29. Mockaitis G, Bruant G, Guiot SR, et al (2020) Acidic and thermal pre-treatments for anaerobic digestion inoculum to improve hydrogen and volatile fatty acid production using xylose as the substrate. Renewable Energy 145: https://doi.org/10.1016/j.renene.2019.06.134

  30. Lide DR, Frederikse HPR (1995) CRC Handbook of Chemistry and Physics, 76th edn. CRC Press Inc., Boca Ratón, FL, USA

    Google Scholar 

  31. Sander R (2015) Compilation of Henry’s law constants (version 4.0) for water as solvent. Atmos Chem Phys 15:4399–4981. https://doi.org/10.5194/acp-15-4399-2015

    Article  Google Scholar 

  32. APHA (2005) Standard methods for the examination of water and wastewater, 21st ed. Washington DC

  33. Dilallo R, Albertson O (1961) Volatile Acids by Direct Titration. J Water Pollut Control Fed 33:356–365

    Google Scholar 

  34. Ripley L, Boyle W, Convrese J (1986) Improved alkalimetric monitoring for anaerobic digestion of high-strength wastes. J Water Pollut Control Fed 58:406–411

    Google Scholar 

  35. Guiot SR, Cimpoia R, Carayon G (2011) Potential of wastewater-treating anaerobic granules for biomethanation of synthesis gas. Environ Sci Technol 45:2006–2012. https://doi.org/10.1021/es102728m

    Article  Google Scholar 

  36. Muyzer G, Dewaal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by desnaturing gradient gel electrophoresis analysis of polymerase chain reaction amplified genes coding for 16S ribosomal RNA. Appl Environ Microbiol 59:695–700

    Article  Google Scholar 

  37. Griffiths RI, Whiteley a S, O’Donnell a G, Bailey MJ, (2000) Rapid method for coextraction of DNA and RNA from natural environments for analysis of ribosomal DNA- and rRNA-based microbial community composition. Appl Environ Microbiol 66:5488–5491

    Article  Google Scholar 

  38. Lévesque MJ, La Boissière S, Thomas JC et al (1997) Rapid method for detecting Desulfitobacterium frappieri strain PCP-1 in soil by the polymerase chain reaction. Appl Microbiol Biotechnol 47:719–725

    Article  Google Scholar 

  39. Berthelet M, Whyte LG, Greer CW (1996) Rapid, direct extraction of DNA from soils for PCR analysis using polyvinylpolypyrrolidone spin columns. FEMS Microbiol Lett 138:17–22

    Article  Google Scholar 

  40. Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73:5261–5267. https://doi.org/10.1128/AEM.00062-07

    Article  Google Scholar 

  41. Claesson MJ, O’Sullivan O, Wang Q et al (2009) Comparative analysis of pyrosequencing and a phylogenetic microarray for exploring microbial community structures in the human distal intestine. PLoS ONE 4:e6669. https://doi.org/10.1371/journal.pone.0006669

    Article  Google Scholar 

  42. Mavrovouniotis ML (1990) Group contributions for estimating standard gibbs energies of formation of biochemical compounds in aqueous solution. Biotechnol Bioeng 36:1070–1082. https://doi.org/10.1002/bit.260361013

    Article  Google Scholar 

  43. Mavrovouniotis ML (1991) Errata Group Contributions for Estimating Standard Gibbs Energies of Formation of Biochemical Compounds in Aqueous Solution 38:803–804

    Google Scholar 

  44. Mockaitis G, Bruant G (2021) Use of hydrogen as electron donor to solventogenesis of volatile organic acids. In: Mendeley Data. https://www.doi.org/10.17632/wwm4zwbgrc

  45. Harper SR, Pohland FG (1985) Recent Developments In Hydrogen Management During Anaerobic Biological Wastewater Treatment. Biotechnol Bioeng 28:585–602

    Article  Google Scholar 

  46. Speece RE (1996) Anaerobic biotechnology for industrial wastewater. Vanderbilt University

  47. Caspi R, Billington R, Fulcher CA et al (2018) The MetaCyc database of metabolic pathways and enzymes. Nucleic Acids Res 46:D633–D639. https://doi.org/10.1093/nar/gkx935

    Article  Google Scholar 

  48. John J, Saranathan R, Adigopula LN et al (2016) The quorum sensing molecule N-acyl homoserine lactone produced by Acinetobacter baumannii displays antibacterial and anticancer properties. Biofouling 32:1029–1047. https://doi.org/10.1080/08927014.2016.1221946

    Article  Google Scholar 

  49. Zhang K, Yang X, Yang J et al (2020) Alcohol dehydrogenase modulates quorum sensing in biofilm formations of Acinetobacter baumannii. Microb Pathog 148:104451. https://doi.org/10.1016/j.micpath.2020.104451

    Article  Google Scholar 

  50. Subhadra B, Hwan OhM, Hee Choi C (2016) Quorum sensing in <em>Acinetobacter</em>: with special emphasis on antibiotic resistance, biofilm formation and quorum quenching. AIMS Microbiology 2:27–41. https://doi.org/10.3934/microbiol.2016.1.27

    Article  Google Scholar 

  51. Grady EN, MacDonald J, Liu L et al (2016) Current knowledge and perspectives of Paenibacillus: A review. Microb Cell Fact 15:203

    Article  Google Scholar 

  52. Leifson E, Hugh R (1954) A new type of polar monotrichous flagellation. J Gen Microbiol 10:68–70. https://doi.org/10.1099/00221287-10-1-68

    Article  Google Scholar 

  53. Caspi R, Altman T, Dale JM et al (2010) The MetaCyc database of metabolic pathways and enzymes and the BioCyc collection of pathway/genome databases. Nucleic Acids Res 38:D473–D479. https://doi.org/10.1093/nar/gkp875

    Article  Google Scholar 

  54. Xu J, Sun L, Xing X et al (2020) Culturing bacteria from fermentation pit muds of Baijiu with culturomics and amplicon-based metagenomic approaches. Front Microbiol 11:1223. https://doi.org/10.3389/fmicb.2020.01223

    Article  Google Scholar 

  55. Tracy BP, Jones SW, Fast AG et al (2012) Clostridia: the importance of their exceptional substrate and metabolite diversity for biofuel and biorefinery applications. Curr Opin Biotechnol 23:364–381. https://doi.org/10.1016/j.copbio.2011.10.008

    Article  Google Scholar 

  56. Ueki A, Hirono T, Sato E et al (1991) Ethanol and amylase production by a newly isolated Clostridium sp. World Journal of Microbiology 7:385–393

    Article  Google Scholar 

  57. Bruant G, Lévesque M-J, Peter C et al (2010) Genomic analysis of carbon monoxide utilization and butanol production by Clostridium carboxidivorans strain P7. PLoS ONE 5:e13033. https://doi.org/10.1371/journal.pone.0013033

    Article  Google Scholar 

Download references

Acknowledgements

The authors thankfully acknowledge Marie-Josée Lévesque, Christine Maynard and Sylvie Sanschagrin for their assistance with the biomolecular techniques (DNA extraction, purification and PCR) and sequencing (through Ion TorrentTM); and also, Stephane Deschamps and Alain Corriveau for their valuable contribution with physicochemical analysis (HPLC and gas chromatography of alcohols).

Funding

This work was supported by FAPESP – Fundação de Amparo a Pesquisa do Estado de São Paulo (processes 2010/18.463–9 and 2013/18.172–2 – G. Mockaitis and 2009/15.984–0 – M. Zaiat), and the NRC – National Research Council of Canada (project A1-004645 – S.R. Guiot).

Author information

Authors and Affiliations

  1. Anaerobic Technologies and Bioprocess Control Group, Energy, Mining and Environment Research Center, National Research Council Canada, 6100 Royalmount Avenue, Montreal, QC, H4P 2R2, Canada

    Gustavo Mockaitis, Guillaume Bruant & Serge R. Guiot

  2. Biological Processes Laboratory, Center for Research, Development and Innovation in Environmental Engineering, São Carlos School of Engineering, University of São Paulo (EESC/USP), Av. João DagnoneSanta Angelina, São Carlos, São Paulo 13563-120, 1100, Brazil

    Eugenio Foresti & Marcelo Zaiat

  3. Interdisciplinary Research Group On Biotechnology Applied To the Agriculture and the Environment, School of Agricultural Engineering, GBMA/FEAGRI/UNICAMP), University of Campinas, CEP, 501 Cândido Rondon Avenue13.083-875, Campinas, SP, Brazil

    Gustavo Mockaitis

Authors
  1. Gustavo Mockaitis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guillaume Bruant
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eugenio Foresti
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marcelo Zaiat
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Serge R. Guiot
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors have planned the experiments. GM performed all the experiments, data treatment, statistics, and mathematical modelling. GB treated and analyzed all sequencing data. All authors have analyzed and discussed the results. GM has written the paper. All authors have read, reviewed, and approved the final manuscript. Experiments were conducted in the Montréal branch of the National Research Council of Canada from January to July of 2014.

Corresponding author

Correspondence to Gustavo Mockaitis.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mockaitis, G., Bruant, G., Foresti, E. et al. Physicochemical pretreatment selects microbial communities to produce alcohols through metabolism of volatile fatty acids. Biomass Conv. Bioref. 14, 2661–2675 (2024). https://doi.org/10.1007/s13399-022-02383-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13399-022-02383-7

Keywords

Search

Navigation