Skip to main content

Effects of Lactobacillus plantarum on the ethanol tolerance of Saccharomyces cerevisiae

Applied Microbiology and Biotechnology volume 105, pages 2597–2611 (2021)Cite this article

Abstract

The bioethanol fermentation by Saccharomyces cerevisiae is often challenged by bacterial contamination, especially lactic acid bacteria (LAB). LAB can inhibit the growth S. cerevisiae by secreting organic acids and competing for nutrients and physical space. However, the range of favorable effects attributed to LAB during bioethanol fermentation, and their associated mechanisms of regulation, are not fully understood. This study was performed to clarify the effects of Lactobacillus plantarum, an important contaminative LAB in bioethanol fermentation, on the mechanism of ethanol tolerance in S. cerevisiae. The results showed that the presence of L. plantarum increased the ethanol tolerance of S. cerevisiae by promoting or inhibiting various metabolic processes in the yeast cells: The metabolism of trehalose, ergosterol, certain amino acids, proton pumps, stress response transcriptional activators, and heat shock proteins were all promoted; amounts of intracellular monounsaturated fatty acids and the accumulation of reactive oxygen species were inhibited. Furthermore, the maintenance of the acquired higher ethanol tolerance of S. cerevisiae was dependent on the coexistence of L. plantarum. These results suggested a complex relationship existed between S. cerevisiae and the contaminating LAB that might also play a beneficial role during fermentation by promoting the ethanol tolerance of yeast. The results from this study suggested that the extent of controlling bacterial contamination on bioethanol fermentation efficiency should be given careful consideration.

Key points

• L. plantarum improved the ethanol tolerance of S. cerevisiae.

• L. plantarum regulated the ethanol tolerance-related metabolism of yeast cells.

• L. plantarum coexistence facilitated maintenance of ethanol tolerance in yeast cells.

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

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

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

Data availability

All data generated or analyzed during this study are included in this published article (and its supplementary information files).

References

  • Abbott DA, Suir E, van Maris AJA, Pronk JT (2008) Physiological and transcriptional responses to high concentrations of lactic acid in anaerobic chemostat cultures of Saccharomyces cerevisiae. Appl Environ Microbiol 74:5759–5768

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Alexandre H, Rousseaux I, Charpentier C (1994) Relationship between ethanol tolerance, lipid composition and plasma membrane fluidity in Saccharomyces cerevisiae and Kloeckera apiculata. FEMS Microbiol Lett 124:17–22

    CAS  PubMed  Article  Google Scholar 

  • Atkinson S, Williams P (2009) Quorum sensing and social networking in the microbial world. J R Soc Interface 6:959–978

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Avbelj M, Zupan J, Kranjc L, Raspor P (2015) Quorum-sensing kinetics in Saccharomyces cerevisiae: a symphony of ARO genes and aromatic alcohols. J Agric Food Chem 63:8544–8550

    CAS  PubMed  Article  Google Scholar 

  • Bailey SM, Pietsch EC, Cunningham CC (1999) Ethanol stimulates the production of reactive oxygen species at mitochondrial complexes I and III. Free Radic Biol Med 27:891–900

    CAS  PubMed  Article  Google Scholar 

  • Benaroudj N, Lee DH, Goldberg AL (2001) Trehalose accumulation during cellular stress protects cells and cellular proteins from damage by oxygen radicals. J Biol Chem 276:24261–24267

    CAS  PubMed  Article  Google Scholar 

  • Bhargava N, Sharma P, Capalash N (2014) Pyocyanin stimulates quorum sensing-mediated tolerance to oxidative stress and increases persister cell populations in Acinetobacter baumannii. Infect Immun 82:3417–3425

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  • Brexó RP, Sant’Ana AS (2017) Impact and significance of microbial contamination during fermentation for bioethanol production. Renew Sust Energ Rev 73:423–434

    Article  CAS  Google Scholar 

  • Brexó RP, Sant’Ana AS (2018) Microbial interactions during sugar cane must fermentation for bioethanol production: does quorum sensing play a role? Crit Rev Biotechnol 38:231–244

    PubMed  Article  CAS  Google Scholar 

  • Cheng Y, Du Z, Zhu H, Guo X, He X (2016) Protective effects of arginine on Saccharomyces cerevisiae against ethanol stress. Sci Rep 6:31311

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Ding J, Huang X, Zhang L, Zhao N, Yang D, Zhang K (2009) Tolerance and stress response to ethanol in the yeast Saccharomyces cerevisiae. Appl Microbiol Biotechnol 85:253–263

    CAS  PubMed  Article  Google Scholar 

  • Divate NR, Chen GH, Wang PM, Ou BR, Chung YC (2016) Engineering Saccharomyces cerevisiae for improvement in ethanol tolerance by accumulation of trehalose. Bioengineered 7:445–458

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Dong SJ, Lin XH, Li H (2015a) Regulation of Lactobacillus plantarum contamination on the carbohydrate and energy related metabolisms of Saccharomyces cerevisiae during bioethanol fermentation. Int J Biochem Cell Biol 68:33–41

    CAS  PubMed  Article  Google Scholar 

  • Dong SJ, Yi CF, Li H (2015b) Changes of Saccharomyces cerevisiae cell membrane components and promotion to ethanol tolerance during the bioethanol fermentation. Int J Biochem Cell Biol 69:196–203

    CAS  PubMed  Article  Google Scholar 

  • Ferreira AK, de Sá-Júnior PL, KFM P, de Azevedo RA, Câmara DA, Costa AS, Figueiredo CR, Matsuo AL, Massaoka MH, Auada AV, Lebrun I, Damião MC, Tavares MT, Magri FM, Kerkis I, Parise Filho R (2014) Cytotoxic effects of dillapiole on MDA-MB-231 cells involve the induction of apoptosis through the mitochondrial pathway by inducing an oxidative stress while altering the cytoskeleton network. Biochimie 99:195–207

    CAS  PubMed  Article  Google Scholar 

  • Gorner W, Durchschlag E, Martinez-Pastor MT, Estruch F, Ammerer G, Hamilton B, Ruis H, Schuller C (1998) Nuclear localization of the C2H2 zinc finger protein Msn2p is regulated by stress and protein kinase A activity. Genes Dev 12:586–597

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Hirasawa T, Yoshikawa K, Nakakura Y, Nagahisa K, Furusawa C, Katakura Y, Shimizu H, Shioya S (2007) Identification of target genes conferring ethanol stress tolerance to Saccharomyces cerevisiae based on DNA microarray data analysis. J Biotechnol 131:34–44

    CAS  PubMed  Article  Google Scholar 

  • Inoue T, Iefuji H, Fujii T, Soga H, Satoh K (2000) Cloning and characterization of a gene complementing the mutation of an ethanol-sensitive mutant of sake yeast. Biosci Biotechnol Biochem 64:229–236

    CAS  PubMed  Article  Google Scholar 

  • Kasavi C, Finore I, Lama L, Nicolaus B, Oliver SG, Toksoy Oner E, Kirdar B (2012) Evaluation of industrial Saccharomyces cerevisiae strains for ethanol production from biomass. Biomass Bioenergy 45:230–238

    CAS  Article  Google Scholar 

  • Li H, Ma ML, Luo S, Zhang RM, Han P, Hu W (2012) Metabolic responses to ethanol in Saccharomyces cerevisiae using a gas chromatography tandem mass spectrometry-based metabolomics approach. Int J Biochem Cell Biol 44:1087–1096

    CAS  PubMed  Article  Google Scholar 

  • Lin X, Han P, Dong S, Li H (2015) Preparation and application of bacteriophage-loaded chitosan microspheres for controlling Lactobacillus plantarum contamination in bioethanol fermentation. RSC Adv 5:69886–69893

    CAS  Article  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Lucena BT, dos Santos BM, Moreira JL, Moreira APB, Nunes AC, Azevedo V, Miyoshi A, Thompson FL, de Morais M (2010) Diversity of lactic acid bacteria of the bioethanol process. BMC Microbiol 10:298

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  • Ma M, Liu ZL (2010) Mechanisms of ethanol tolerance in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 87:829–845

    CAS  PubMed  Article  Google Scholar 

  • Morita Y, Nakamori S, Takagi H (2002) Effect of proline and arginine metabolism on freezing stress of Saccharomyces cerevisiae. J Biosci Bioeng 94:390–394

    CAS  PubMed  Article  Google Scholar 

  • Muthaiyan A, Limayem A, Ricke SC (2011) Antimicrobial strategies for limiting bacterial contaminants in fuel bioethanol fermentations. Prog Energy Combust 37:351–370

    CAS  Article  Google Scholar 

  • Narendranath NV, Hynes SH, Thomas KC, Ingledew WM (1997) Effects of Lactobacilli on yeast-catalyzed ethanol fermentations. Appl Environ Microbiol 63:4158–4163

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Rich JO, Leathers TD, Bischoff KM, Anderson AM, Nunnally MS (2015) Biofilm formation and ethanol inhibition by bacterial contaminants of biofuel fermentation. Bioresour Technol 196:347–354

    CAS  PubMed  Article  Google Scholar 

  • Rosa MF, Sá-Correia I (1992) Ethanol tolerance and activity of plasma membrane ATPase in Kluyveromyces marxianus and Saccharomyces cerevisiae. Enzyme Microb Technol 14:23–27

    CAS  Article  Google Scholar 

  • Rossouw D, Du Toit M, Bauer FF (2012) The impact of co-inoculation with Oenococcus oeni on the trancriptome of Saccharomyces cerevisiae and on the flavour-active metabolite profiles during fermentation in synthetic must. Food Microbiol 29:121–131

    CAS  PubMed  Article  Google Scholar 

  • Rubin EM (2008) Genomics of cellulosic biofuels. Nature 454:841–845

    CAS  PubMed  Article  Google Scholar 

  • Saini P, Beniwal A, Kokkiligadda A, Vij S (2018) Response and tolerance of yeast to changing environmental stress during ethanol fermentation. Process Biochem 72:1–12

    CAS  Article  Google Scholar 

  • Shobayashi M, Mitsueda S, Ago M, Fujii T, Iwashita K, Iefuji H (2005) Effects of culture conditions on ergosterol biosynthesis by Saccharomyces cerevisiae. Biosci Biotechnol Biochem 69:2381–2388

    CAS  PubMed  Article  Google Scholar 

  • Sieuwerts S, Bron PA, Smid EJ (2018) Mutually stimulating interactions between lactic acid bacteria and Saccharomyces cerevisiae in sourdough fermentation. LWT-Food Sci Technol 90:201–206

    CAS  Article  Google Scholar 

  • Szeto SS, Reinke SN, Sykes BD, Lemire BD (2010) Mutations in the Saccharomyces cerevisiae succinate dehydrogenase result in distinct metabolic phenotypes revealed through 1H NMR-based metabolic footprinting. J Proteome Res 9:6729–6739

    CAS  PubMed  Article  Google Scholar 

  • Takagi H, Takaoka M, Kawaguchi A, Kubo Y (2005) Effect of L-proline on sake brewing and ethanol stress in Saccharomyces cerevisiae. Appl Environ Microbiol 71:8656–8662

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Takagi H, Taguchi J, Kaino T (2016) Proline accumulation protects Saccharomyces cerevisiae cells in stationary phase from ethanol stress by reducing reactive oxygen species levels: Proline protects yeast cells from ethanol by reducing ROS levels. Yeast 33:355–363

    CAS  PubMed  Article  Google Scholar 

  • Teparić R, Stuparević I, Mrsa V (2004) Increased mortality of Saccharomyces cerevisiae cell wall protein mutants. Microbiology 150:3145–3150

    PubMed  Article  CAS  Google Scholar 

  • Thomas KC, Hynes SH, Ingledew WM (2001) Effect of lactobacilli on yeast growth, viability and batch and semi-continuous alcoholic fermentation of corn mash. J Appl Microbiol 90:819–828

    CAS  PubMed  Article  Google Scholar 

  • Tian J, Ji Z, Wang F, Song M, Li H (2017) The toxic effects of tetrachlorobisphenol a in Saccharomyces cerevisiae cells via metabolic interference. Sci Rep 7:2655

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  • Vamvakas S, Kapolos J, Farmakis L, Genneos F, Damianaki M, Chouli X, Vardakou A, Liosi S, Stavropoulou E, Leivaditi E, Fragki M, Labrakou E, Gashi E, Demoli D (2019) Specific serine residues of Msn2/4 are responsible for regulation of alcohol fermentation rates and ethanol resistance. Biotechnol Prog 35:e2759

    PubMed  Article  CAS  Google Scholar 

  • van Voorst F, Houghton-Larsen J, Jønson L, Kielland-Brandt MC, Brandt A (2006) Genome-wide identification of genes required for growth of Saccharomyces cerevisiae under ethanol stress. Yeast 23:351–359

    PubMed  Article  CAS  Google Scholar 

  • Yi C, Wang F, Dong S, Li H (2016) Changes of trehalose content and expression of relative genes during the bioethanol fermentation by Saccharomyces cerevisiae. Can J Microbiol 62:827–835

    CAS  PubMed  Article  Google Scholar 

  • Zhang M, Shi J, Jiang L (2015) Modulation of mitochondrial membrane integrity and ROS formation by high temperature in Saccharomyces cerevisiae. Electron J Biotechnol 18:202–209

    Article  Google Scholar 

Download references

Funding

This work was supported by the National Key Research and Development Program of China (No. 2018YFA0903000), the National Science and Technology Major Project during the 13th 5-Year Plan Period (No. 2019ZX09721001-007-002), Shenzhen Science and Technology Project (No. JCYJ20180507183842516), and the Higher Education and High-quality and World-Class Universities (No. PY201617).

Author information

Author notes

  1. Xianlin He, Bo Liu and Yali Xu contributed equally to this work.

Affiliations

  1. Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China

    Xianlin He, Yali Xu, Ze Chen & Hao Li

  2. Shandong Provincial Key Laboratory of Biopharmaceuticals, Shandong Academy of Pharmaceutical Sciences, Jinan, 250101, China

    Bo Liu

Authors
  1. Xianlin He
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bo Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yali Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ze Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hao Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

XH and HL conceived and designed research. XH, BL, and YX conducted experiments. XH and ZC analyzed data. XH, BL, YX, and HL wrote the manuscript. All authors read and approved the manuscript.

Corresponding author

Correspondence to Hao Li.

Ethics declarations

Ethics approval

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

Conflict of interest

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.

Supplementary Information

ESM 1

(PDF 566 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

He, X., Liu, B., Xu, Y. et al. Effects of Lactobacillus plantarum on the ethanol tolerance of Saccharomyces cerevisiae. Appl Microbiol Biotechnol 105, 2597–2611 (2021). https://doi.org/10.1007/s00253-021-11198-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-021-11198-x

Keywords

  • Saccharomyces cerevisiae
  • Lactobacillus plantarum
  • Bacterial contamination
  • Regulation of ethanol tolerance
  • Metabolism