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Applied Microbiology and Biotechnology

, Volume 103, Issue 12, pp 5039–5050 | Cite as

Effect of contamination with Lactobacillus fermentum I2 on ethanol production by Spathaspora passalidarum

  • Karen Cristina Collograi
  • Aline Carvalho da Costa
  • Jaciane Lutz IenczakEmail author
Bioenergy and biofuels
  • 86 Downloads

Abstract

Second-generation bioethanol is a promising source of renewable energy. In Brazilian mills, the production of ethanol from sugarcane (first generation, 1G) is a consolidated process performed by Saccharomyces cerevisiae and characterized by high substrate concentrations, high cell density, and cell recycle. The main bacterial contaminants in 1G fermentation tanks are lactic acid bacteria, especially bacteria from the Lactobacillus genus, which is associated with a decrease in ethanol yield and yeast cell viability, among other negative effects. Second-generation (2G) bioethanol production is characterized by the conversion of glucose and xylose into ethanol by genetically modified or non-Saccharomyces yeasts. Spathaspora passalidarum is a promising non-Saccharomyces yeast for 2G ethanol production due to its ability to effectively convert xylose into ethanol. The effect of bacterial contamination on the fermentation of this yeast is unknown; therefore, L. fermentum, a common bacterium found in Brazilian 1G processes, was studied in coculture with S. passalidarum in a fed-batch fermentation process similar to that used in 1G mills. Individually, L. fermentum I2 was able to simultaneously consume glucose and xylose in nutrient-rich broth (Man, Rogosa, and Sharpe (MRS + xylose) but failed to grow in a glucose- and xylose-based synthetic broth. In coculture with S. passalidarum, the bacteria remained at a concentration of 108 UFC/mL throughout cell recycling, but no flocculation was observed, and it did not affect the fermentative parameters or the cellular viability of the yeast. Under both conditions, the maximum ethanol production was 21 g L−1 with volumetric productivity ranging from 0.65 to 0.70 g L−1 h−1. S. passalidarum was thus shown to be resistant to L. fermentum I2 under the conditions studied.

Keywords

Second-generation ethanol Microbial contamination Non-conventional yeast recycle 

Notes

Acknowledgments

We are grateful to the CNPEM and CTBE for assigning the structures for the realization of the experiments and for the help in HPLC analysis.

Funding information

The authors acknowledge the financial assistance of the FAPESP- São Paulo Research Foundation (processes no 2017/04997-0 and 2016/06142-0) and the CNPQ - National Council for Scientific and Technological Development (process no 132142/2017-1).

Compliance with ethical standards

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

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Albers E, Johansson E, Franzén CJ, Larsson C (2011) Selective suppression of bacterial contaminants by process conditions during lignocellulose based yeast fermentations. Biotechnol Biofuels 4:59.  https://doi.org/10.1186/1754-6834-4-59 CrossRefGoogle Scholar
  2. Alfenore S, Molina-Jouve C, Guillouet SE, Uribelarrea JL, Goma G, Benbadis L (2002) Improving ethanol production and viability of Saccharomyces cerevisiae by a vitamin feeding strategy during fed-batch process. Appl Microbiol Biotechnol 60:67–72.  https://doi.org/10.1007/s00253-002-1092-7 CrossRefGoogle Scholar
  3. Amorim HV, Lopes ML, De Castro Oliveira JV, Buckeridge MS, Goldman GH (2011) Scientific challenges of bioethanol production in Brazil. Appl Microbiol Biotechnol 91:1267–1275.  https://doi.org/10.1007/s00253-011-3437-6 CrossRefGoogle Scholar
  4. Araújo TM, Souza MT, Diniz RHS, Yamakawa CK, Soares LB, Lenczak JL, de Castro Oliveira JV, Goldman GH, Barbosa EA, Campos ACS, Castro IM, Brandão RL (2018) Cachaça yeast strains: alternative starters to produce beer and bioethanol. Antonie Van Leeuwenhoek, Int J Gen Mol Microbiol 111:1749–1766.  https://doi.org/10.1007/s10482-018-1063-3 CrossRefGoogle Scholar
  5. Bassi APG, Meneguello L, Paraluppi AL, Sanches BCP, Ceccato-Antonini SR (2018) Interaction of Saccharomyces cerevisiaeLactobacillus fermentumDekkera bruxellensis and feedstock on fuel ethanol fermentation. Antonie Van Leeuwenhoek, Int J Gen Mol Microbiol 111:1661–1672.  https://doi.org/10.1007/s10482-018-1056-2
  6. Basso LC, de Amorim HV, de Oliveira AJ, Lopes ML (2008) Yeast selection for fuel ethanol production in Brazil. FEMS Yeast Res 8:1155–1163.  https://doi.org/10.1111/j.1567-1364.2008.00428.x CrossRefGoogle Scholar
  7. Basso TO, Gomes FS, Lopes ML, De Amorim HV, Eggleston G, Basso LC (2014) Homo- and heterofermentative lactobacilli differently affect sugarcane-based fuel ethanol fermentation. Antonie Van Leeuwenhoek, Int J Gen Mol Microbiol 105:169–177.  https://doi.org/10.1007/s10482-013-0063-6 CrossRefGoogle Scholar
  8. Bonatelli ML, Quecine MC, Silva MS, Labate CA (2017) Characterization of the contaminant bacterial communities in sugarcane first-generation industrial ethanol production. FEMS Microbiol Lett 364:1–8.  https://doi.org/10.1093/femsle/fnx159 CrossRefGoogle Scholar
  9. Brandenburg J, Poppele I, Blomqvist J, Puke M, Pickova J, Sandgren M, Rapoport A, Vedernikovs N, Passoth V (2018) Bioethanol and lipid production from the enzymatic hydrolysate of wheat straw after furfural extraction. Appl Microbiol Biotechnol 102:6269–6277.  https://doi.org/10.1007/s00253-018-9081-7 CrossRefGoogle Scholar
  10. Brexó RP, Sant’Ana AS (2017) Impact and significance of microbial contamination during fermentation for bioethanol production. Renew Sust Energ Rev 73:423–434.  https://doi.org/10.1016/j.rser.2017.01.151 CrossRefGoogle Scholar
  11. Carpio LGT, Simone de Souza F (2017) Optimal allocation of sugarcane bagasse for producing bioelectricity and second generation ethanol in Brazil: scenarios of cost reductions. Renew Energy 111:771–780.  https://doi.org/10.1016/j.renene.2017.05.015 CrossRefGoogle Scholar
  12. Carvalho-Netto OV, Carazzolle MF, Mofatto LS, Teixeira PJPL, Noronha MF, Calderón LAL, Mieczkowski PA, Argueso LL, Pereira GAG (2015) Saccharomyces cerevisiae transcriptional reprograming due to bacterial contamination during industrial scale bioethanol production. Microb Cell Factories 14:1–13.  https://doi.org/10.1186/s12934-015-0196-6 CrossRefGoogle Scholar
  13. Costa MAS, Cerri BC, Ceccato-Antonini SR (2018) Ethanol addition enhances acid treatment to eliminate Lactobacillus fermentum from the fermentation process for fuel ethanol production. Lett Appl Microbiol 66:77–85.  https://doi.org/10.1111/lam.12819 CrossRefGoogle Scholar
  14. de Carvalho DM, de Queiroz JH, Colodette JL (2016) Assessment of alkaline pretreatment for the production of bioethanol from eucalyptus, sugarcane bagasse and sugarcane straw. Ind Crop Prod 94:932–941.  https://doi.org/10.1016/j.indcrop.2016.09.069 CrossRefGoogle Scholar
  15. Fitzpatrick JJ, O’Keeffe U (2001) Influence of whey protein hydrolysate addition to whey permeate batch fermentations for producing lactic acid. Process Biochem 37:183–186.  https://doi.org/10.1016/S0032-9592(01)00203-5 CrossRefGoogle Scholar
  16. Gombert AK, van Maris AJA (2015) Improving conversion yield of fermentable sugars into fuel ethanol in 1st generation yeast-based production processes. Curr Opin Biotechnol 33:81–86.  https://doi.org/10.1016/j.copbio.2014.12.012 CrossRefGoogle Scholar
  17. Hahn-Hägerdal B, Karhumaa K, Fonseca C, Spencer-Martins I, Gorwa-Grauslund MF (2007) Towards industrial pentose-fermenting yeast strains. Appl Microbiol Biotechnol 74:937–953.  https://doi.org/10.1007/s00253-006-0827-2 CrossRefGoogle Scholar
  18. Hou X, Yao S (2012) Improved inhibitor tolerance in xylose-fermenting yeast Spathaspora passalidarum by mutagenesis and protoplast fusion. Appl Microbiol Biotechnol 93:2591–2601.  https://doi.org/10.1007/s00253-011-3693-5 CrossRefGoogle Scholar
  19. Jönsson LJ, Martín C (2016) Pretreatment of lignocellulose: formation of inhibitory by-products and strategies for minimizing their effects. Bioresour Technol 199:103–112.  https://doi.org/10.1016/j.biortech.2015.10.009 CrossRefGoogle Scholar
  20. Long TM, Su YK, Headman J, Higbee A, Willis LB, Jeffries TW (2012) Cofermentation of glucose, xylose, and cellobiose by the beetle-associated yeast Spathaspora passalidarum. Appl Environ Microbiol 78:5492–5500.  https://doi.org/10.1128/AEM.00374-12 CrossRefGoogle Scholar
  21. Lopes ML, Paulillo SC de L, Godoy A, Cherubin RA, Lorenzi MS, Giometti FHC, Bernardino CD, de Amorim Neto HB, de Amorim HV (2016) Ethanol production in Brazil: a bridge between science and industry. Braz J Microbiol 47:64–76.  https://doi.org/10.1016/j.bjm.2016.10.003 CrossRefGoogle Scholar
  22. Lucena BTL, Santos BM, Moreira JLS, Moreira APB, Nunes AC, Azevedo V, Miyoshi A, Thompson FL, Antonio M, Junior DM (2010) Diversity of lactic acid bacteria of the bioethanol process. BMC Microbiol 10:298.  https://doi.org/10.1186/1471-2180-10-298 CrossRefGoogle Scholar
  23. Macrelli S, Mogensen J, Zacchi G (2012) Techno economic evaluation of 2nd generation bioethanol production from sugar cane bagasse and leaves integrated with the sugar based ethanol process. Biotechnol Biofuels 5:22.  https://doi.org/10.1186/1754-6834-5-22 CrossRefGoogle Scholar
  24. Muthaiyan A, Limayem A, Ricke SC (2011) Antimicrobial strategies for limiting bacterial contaminants in fuel bioethanol fermentations. Prog Energy Combust Sci 37:351–370.  https://doi.org/10.1016/j.pecs.2010.06.005 CrossRefGoogle Scholar
  25. Nakanishi SC, Soares LB, Biazi LE, Nascimento VM, Costa AC, Rocha GJM, Ienczak JL (2017) Fermentation strategy for second generation ethanol production from sugarcane bagasse hydrolyzate by Spathaspora passalidarum and Scheffersomyces stipitis. Biotechnol Bioeng 9999:1–11.  https://doi.org/10.1002/bit.26357
  26. Palmqvist E, Hahn-Hägerdal B (2000) Fermentation of lignocellulosic hydrolysates. II: inhibitors and mechanisms of inhibition. Bioresour Technol 74:25–33.  https://doi.org/10.1016/S0960-8524(99)00161-3 CrossRefGoogle Scholar
  27. Paulova L, Patakova P, Branska B, Rychtera M, Melzoch K (2015) Lignocellulosic ethanol: technology design and its impact on process efficiency. Biotechnol Adv 33:1091–1107.  https://doi.org/10.1016/j.biotechadv.2014.12.002 CrossRefGoogle Scholar
  28. Reis VR, Bassi APG, Cerri BC, Almeida AR, Carvalho IGB, Bastos RG, Ceccato-Antonini SR (2018) Effects of feedstock and co-culture of Lactobacillus fermentum and wild Saccharomyces cerevisiae strain during fuel ethanol fermentation by the industrial yeast strain PE-2. AMB Express 8:23.  https://doi.org/10.1186/s13568-018-0556-9 CrossRefGoogle Scholar
  29. Rocha GJ d M, Nascimento VM, Gonçalves AR, Silva VFN, Martín C (2015) Influence of mixed sugarcane bagasse samples evaluated by elemental and physical-chemical composition. Ind Crop Prod 64:52–58.  https://doi.org/10.1016/j.indcrop.2014.11.003 CrossRefGoogle Scholar
  30. Santos SC, de Sousa AS, Dionisio SR, Tramontina R, Ruller R, Squina FM, Vaz Rossell CE, da Costa AC, Ienczak JL (2016) Bioethanol production by recycled Scheffersomyces stipitis in sequential batch fermentations with high cell density using xylose and glucose mixture. Bioresour Technol 219:319–329.  https://doi.org/10.1016/j.biortech.2016.07.102 CrossRefGoogle Scholar
  31. Schell DJ, Dowe N, Ibsen KN, Riley CJ, Ruth MF, Lumpkin RE (2007) Contaminant occurrence , identification and control in a pilot-scale corn fiber to ethanol conversion process. 98:2942–2948.  https://doi.org/10.1016/j.biortech.2006.10.002
  32. Su YK, Willis LB, Jeffries TW (2015) Effects of aeration on growth, ethanol and polyol accumulation by Spathaspora passalidarum NRRL Y-27907 and Scheffersomyces stipitis NRRL Y-7124. Biotechnol Bioeng 112:457–469.  https://doi.org/10.1002/bit.25445 CrossRefGoogle Scholar
  33. Veras HCT, Parachin NS, Almeida JRM (2017) Comparative assessment of fermentative capacity of different xylose-consuming yeasts. Microb Cell Factories 16:1–8.  https://doi.org/10.1186/s12934-017-0766-x CrossRefGoogle Scholar
  34. Zhang C, Guo T, Xin Y, Gao X, Kong J (2016) Catabolite responsive element deficiency of xyl operon resulting in carbon catabolite derepression in Lactobacillus fermentum 1001. J Appl Microbiol 120:126–137.  https://doi.org/10.1111/jam.12990 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Brazilian Bioethanol Science and Technology Laboratory - CTBE/CNPEMCampinasBrazil
  2. 2.School of Chemical EngineeringState University of Campinas – UNICAMPCampinasBrazil
  3. 3.Chemical Engineering and Food Engineering DepartmentSanta Catarina Federal UniversityFlorianópolisBrazil

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