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Biotechnology Letters

, Volume 41, Issue 11, pp 1309–1318 | Cite as

Evidence for chaotropicity/kosmotropicity offset in a yeast growth model

  • Joshua Eardley
  • Cinzia Dedi
  • Marcus Dymond
  • John E. Hallsworth
  • David J. TimsonEmail author
Original Research Paper

Abstract

Chaotropes are compounds which cause the disordering, unfolding and denaturation of biological macromolecules. It is the chaotropicity of fermentation products that often acts as the primary limiting factor in ethanol and butanol fermentations. Since ethanol is mildly chaotropic at low concentrations, it prevents the growth of the producing microbes via its impacts on a variety of macromolecular systems and their functions. Kosmotropes have the opposite effect to chaotropes and we hypothesised that it might be possible to use these to mitigate chaotrope-induced inhibition of Saccharomyces cerevisiae growth. We also postulated that kosmotrope-mediated mitigation of chaotropicity is not quantitatively predictable. The chaotropes ethanol and urea, and compatible solutes glycerol and betaine (kosmotrope), and the highly kosmotropic salt ammonium sulphate all inhibited the growth rate of Saccharomyces cerevisiae in the concentration range 5–15%. They resulted in increased lag times, decreased maximum specific growth rates, and decreased final optical densities. Surprisingly, neither the stress protectants nor ammonium sulphate reduced the inhibition of growth caused by ethanol. Whereas, in some cases, compatible solutes and kosmotropes mitigated against the inhibitory effects of urea. However, this effect was not mathematically additive from the quantification of chao-/kosmotropicity of each individual compound. The potential effects of glycerol, betaine and/or ammonium sulphate may have been reduced or masked by the metabolic production of compatible solutes. It may nevertheless be that the addition of kosmotropes to fermentations which produce chaotropic products can enhance metabolic activity, growth rate, and/or product formation.

Keywords

Entropy Biofuel Saccharomyces cerevisiae Urea Glycerol Ammonium sulphate 

Notes

Acknowledgements

JE is in receipt of a PhD studentship from the University of Brighton under the Universities Alliance Doctoral Training Alliance in Energy. We thank Dr Lucas Bowler for advice on yeast growth monitoring using the Ascent iEMS Multiskan microplate reader.

Supporting information

Supplementary Figure S1—Effects of chaotropes, kosmotropes and compatible solutes on the growth of S. cerevisiae NCYC 1088. These growth curves were used to derive the growth parameters shown in Figures 1 and 2.

Supplementary Figure S2—Effects of kosmotropes and compatible solutes on the growth of S. cerevisiae NCYC 1088 when challenged with ethanol or urea. These data were used to obtain the growth parameters shown in Figures 3 and 4.

Author contributions

DJT conceived the project and drafted the manuscript, which was co-authored by JE, CD, MD and JEH. JE carried out the majority of the experimental work, assisted by CD and supervised by MD and DJT. JEH provided intellectual input and challenge.

Supplementary material

10529_2019_2737_MOESM1_ESM.tif (294 kb)
Electronic supplementary material 1 (TIF 294 kb)
10529_2019_2737_MOESM2_ESM.tif (333 kb)
Electronic supplementary material 2 (TIF 333 kb)

References

  1. Aviram I (1973) The interaction of chaotropic anions with acid ferricytochrome c. J Biol Chem 248:1894–1896PubMedGoogle Scholar
  2. Ball P, Hallsworth JE (2015) Water structure and chaotropicity: their uses, abuses and biological implications. Phys Chem Chem Phys 17:8297–8305.  https://doi.org/10.1039/c4cp04564e CrossRefPubMedGoogle Scholar
  3. Bell AN, Magill E, Hallsworth JE, Timson DJ (2013) Effects of alcohols and compatible solutes on the activity of β-galactosidase. Appl Biochem Biotechnol 169:786–794.  https://doi.org/10.1007/s12010-012-0003-3 CrossRefPubMedGoogle Scholar
  4. Bennion BJ, Daggett V (2003) The molecular basis for the chemical denaturation of proteins by urea. Proc Natl Acad Sci USA 100:5142–5147.  https://doi.org/10.1073/pnas.0930122100 CrossRefPubMedGoogle Scholar
  5. Bevilacqua A, Speranza B, Sinigaglia M, Corbo MR (2015) A focus on the death kinetics in predictive microbiology: benefits and limits of the most important models and some tools dealing with their application in foods. Foods (Basel, Switzerland) 4:565–580.  https://doi.org/10.3390/foods4040565 CrossRefGoogle Scholar
  6. Bhaganna P et al (2010) Hydrophobic substances induce water stress in microbial cells. Microb Biotechnol 3:701–716.  https://doi.org/10.1111/j.1751-7915.2010.00203.x CrossRefPubMedPubMedCentralGoogle Scholar
  7. Bhaganna P, Bielecka A, Molinari G, Hallsworth JE (2016) Protective role of glycerol against benzene stress: insights from the Pseudomonas putida proteome. Curr Genet 62:419–429.  https://doi.org/10.1007/s00294-015-0539-1 CrossRefPubMedGoogle Scholar
  8. Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM, van der Noordaa J (1990) Rapid and simple method for purification of nucleic acids. J Clin Microbiol 28:495–503PubMedPubMedCentralGoogle Scholar
  9. Brown AD (1978) Compatible solutes and extreme water stress in eukaryotic micro-organisms. Adv Microb Physiol 17:181–242CrossRefGoogle Scholar
  10. Brown AD (1990) Microbial water stress physiology vol First. Vol Book, Whole. Wiley, Chichester, UKGoogle Scholar
  11. Brown AD, Simpson JR (1972) Water relations of sugar-tolerant yeasts: the role of intracellular polyols. J Gen Microbiol 72:589–591CrossRefGoogle Scholar
  12. Chin JP et al (2010) Solutes determine the temperature windows for microbial survival and growth. Proc Natl Acad Sci USA 107:7835–7840.  https://doi.org/10.1073/pnas.1000557107 CrossRefPubMedGoogle Scholar
  13. Coroller L, Leguerinel I, Mettler E, Savy N, Mafart P (2006) General model, based on two mixed weibull distributions of bacterial resistance, for describing various shapes of inactivation curves. Appl Environ Microbiol 72:6493–6502.  https://doi.org/10.1128/aem.00876-06 CrossRefPubMedPubMedCentralGoogle Scholar
  14. Cray JA, Russell JT, Timson DJ, Singhal RS, Hallsworth JE (2013) A universal measure of chaotropicity and kosmotropicity. Environ Microbiol 15:287–296.  https://doi.org/10.1111/1462-2920.12018 CrossRefPubMedGoogle Scholar
  15. Cray JA et al (2015) Chaotropicity: a key factor in product tolerance of biofuel-producing microorganisms. Curr Opin Biotechnol 33:228–259.  https://doi.org/10.1016/j.copbio.2015.02.010 CrossRefPubMedGoogle Scholar
  16. Cray JA, Connor MC, Stevenson A, Houghton JD, Rangel DE, Cooke LR, Hallsworth JE (2016) Biocontrol agents promote growth of potato pathogens, depending on environmental conditions. Microb Biotechnol 9:330–354.  https://doi.org/10.1111/1751-7915.12349 CrossRefPubMedPubMedCentralGoogle Scholar
  17. Das A, Mukhopadhyay C (2009) Urea-mediated protein denaturation: a consensus view. J Phys Chem B 113:12816–12824.  https://doi.org/10.1021/jp906350s CrossRefPubMedGoogle Scholar
  18. de Lima Alves F et al (2015) Concomitant osmotic and chaotropicity-induced stresses in Aspergillus wentii: compatible solutes determine the biotic window. Curr Genet 61:457–477.  https://doi.org/10.1007/s00294-015-0496-8 CrossRefGoogle Scholar
  19. Fox-Powell MG, Hallsworth JE, Cousins CR, Cockell CS (2016) Ionic strength is a barrier to the habitability of mars. Astrobiology 16:427–442.  https://doi.org/10.1089/ast.2015.1432 CrossRefPubMedGoogle Scholar
  20. Gompertz B (1825) On the nature of the function expressive of the law of human mortality, and on a new mode of determining the value of life contingencies. Phil Trans R Soc 115:513–585CrossRefGoogle Scholar
  21. Hallsworth JE (1998) Ethanol-induced water stress in yeast. J Ferment Bioeng 85:125–137.  https://doi.org/10.1016/S0922-338X(97)86756-6 CrossRefGoogle Scholar
  22. Hallsworth JE, Prior BA, Nomura Y, Iwahara M, Timmis KN (2003) Compatible solutes protect against chaotrope (ethanol)-induced, nonosmotic water stress. Appl Environ Microbiol 69:7032–7034CrossRefGoogle Scholar
  23. Hallsworth JE et al (2007) Limits of life in MgCl2-containing environments: chaotropicity defines the window. Environ Microbiol 9:801–813.  https://doi.org/10.1111/j.1462-2920.2006.01212.x CrossRefPubMedGoogle Scholar
  24. Hatefi Y, Hanstein WG (1969) Solubilization of particulate proteins and nonelectrolytes by chaotropic agents. Proc Natl Acad Sci USA 62:1129–1136CrossRefGoogle Scholar
  25. Kella NK, Kinsella JE (1988) Structural stability of β-lactoglobulin in the presence of kosmotropic salts. A kinetic and thermodynamic study. Int J Pept Protein Res 32:396–405CrossRefGoogle Scholar
  26. Krebs HA (1942) Urea formation in mammalian liver. Biochem J 36:758–767CrossRefGoogle Scholar
  27. Kresheck G, Benjamin L (1964) Calorimetric studies of the hydrophobic nature of several protein constituents and ovalbumin in water and in aqueous urea. J Phys Chem 68:2476–2486CrossRefGoogle Scholar
  28. La Cono V et al (2019) The discovery of Lake Hephaestus, the youngest athalassohaline deep-sea formation on Earth. Sci Rep 9:1679.  https://doi.org/10.1038/s41598-018-38444-z CrossRefPubMedPubMedCentralGoogle Scholar
  29. Miyawaki O, Tatsuno M (2011) Thermodynamic analysis of alcohol effect on thermal stability of proteins. J Biosci Bioeng 111:198–203.  https://doi.org/10.1016/j.jbiosc.2010.09.007 CrossRefPubMedGoogle Scholar
  30. Moelbert S, Normand B, De Los Rios P (2004) Kosmotropes and chaotropes: modelling preferential exclusion, binding and aggregate stability. Biophys Chem 112:45–57.  https://doi.org/10.1016/j.bpc.2004.06.012 CrossRefPubMedGoogle Scholar
  31. NCYC (2019) NCYC 1088. National Centre for Yeast Cultures. https://www.ncyc.co.uk/catalogue/saccharomyces-cerevisiae-1088. Accessed 2 Sep 2019
  32. Pace CN (1986) Determination and analysis of urea and guanidine hydrochloride denaturation curves. Methods Enzymol 131:266–280CrossRefGoogle Scholar
  33. Rupley J (1964) The effect of urea and amides upon water structure. J Phys Chem 68:2002–2003CrossRefGoogle Scholar
  34. Salvi G, De Los Rios P, Vendruscolo M (2005) Effective interactions between chaotropic agents and proteins. Proteins 61:492–499.  https://doi.org/10.1002/prot.20626 CrossRefPubMedGoogle Scholar
  35. Stevenson A et al (2015) Is there a common water-activity limit for the three domains of life? ISME J 9:1333–1351.  https://doi.org/10.1038/ismej.2014.219 CrossRefPubMedGoogle Scholar
  36. Stevenson A et al (2017) Glycerol enhances fungal germination at the water-activity limit for life. Environ Microbiol 19:947–967.  https://doi.org/10.1111/1462-2920.13530 CrossRefPubMedGoogle Scholar
  37. Vagenende V, Yap MG, Trout BL (2009) Mechanisms of protein stabilization and prevention of protein aggregation by glycerol. Biochemistry 48:11084–11096.  https://doi.org/10.1021/bi900649t CrossRefPubMedGoogle Scholar
  38. Van Ness J, Chen L (1991) The use of oligodeoxynucleotide probes in chaotrope-based hybridization solutions. Nucleic Acids Res 19:5143–5151CrossRefGoogle Scholar
  39. Williams JP, Hallsworth JE (2009) Limits of life in hostile environments: no barriers to biosphere function? Environ Microbiol 11:3292–3308.  https://doi.org/10.1111/j.1462-2920.2009.02079.x CrossRefPubMedPubMedCentralGoogle Scholar
  40. Wingfield P (1998) Protein precipitation using ammonium sulfate. Curr Protoc Prot Sci 13:A.3F.1–A.3F.8CrossRefGoogle Scholar
  41. Yakimov MM et al (2015) Microbial community of the deep-sea brine Lake Kryos seawater-brine interface is active below the chaotropicity limit of life as revealed by recovery of mRNA. Environ Microbiol 17:364–382.  https://doi.org/10.1111/1462-2920.12587 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Joshua Eardley
    • 1
  • Cinzia Dedi
    • 1
  • Marcus Dymond
    • 1
  • John E. Hallsworth
    • 2
  • David J. Timson
    • 1
    Email author
  1. 1.School of Pharmacy and Biomolecular SciencesUniversity of BrightonBrightonUK
  2. 2.School of Biological Sciences and Institute for Global Food SecurityQueen’s University BelfastBelfastUK

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