Abstract
During alcoholic fermentation, the products build up and can, ultimately, kill the organism due to their effects on the cell’s macromolecular systems. The effects of alcohols on the steady-state kinetic parameters of the model enzyme β-galactosidase were studied. At modest concentrations (0 to 2 M), there was little effect of methanol, ethanol, propanol and butanol on the kinetic constants. However, above these concentrations, each alcohol caused the maximal rate, V max, to fall and the Michaelis constant, K m, to rise. Except in the case of methanol, the chaotropicity of the solute, rather than its precise chemical structure, determined and can, therefore, be used to predict inhibitory activity. Compounds which act as compatible solutes (e.g. glycerol and other polyols) generally reduced enzyme activity in the absence of alcohols at the concentration tested (191 mM). In the case of the ethanol- or propanol-inhibited β-galactosidase, the addition of compatible solutes was unable to restore the enzyme’s kinetic parameters to their uninhibited levels; addition of chaotropic solutes such as urea tended to enhance the effects of these alcohols. It is possible that the compatible solutes caused excessive rigidification of the enzyme’s structure, whereas the alcohols disrupt the tertiary and quaternary structure of the protein. From the point of view of protecting enzyme activity, it may be unwise to add compatible solutes in the early stages of industrial fermentations; however, there may be benefits as the alcohol concentration increases.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Hallsworth, J. E. (1998). Journal of Fermentation and Bioengineering, 85, 125–137.
Hallsworth, J. E., Heim, S., & Timmis, K. N. (2003). Environmental Microbiology, 5, 1270–1280.
Hallsworth, J. E., Prior, B. A., Nomura, Y., Iwahara, M., & Timmis, K. N. (2003). Applied and Environmental Microbiology, 69, 7032–7034.
Bhaganna, P., Volkers, R. J., Bell, A. N., Kluge, K., Timson, D. J., McGrath, J. W., et al. (2010). Microbial Biotechnology, 3, 701–716.
Nicolaou, S. A., Gaida, S. M., & Papoutsakis, E. T. (2010). Metabolic Engineering, 12, 307–331.
Shifrin, S., & Hunn, G. (1969). Archives of Biochemistry and Biophysics, 130, 530–535.
Matsue, S., & Miyawaki, O. (2000). Enzyme MicrobTechnol, 26, 342–347.
Pereira-Rodriguez, A., Fernandez-Leiro, R., Gonzalez-Siso, M. I., Cerdan, M. E., Becerra, M., & Sanz-Aparicio, J. (2012). Journal of Structural Biology, 177, 392–401.
Dickson, R. C., Dickson, L. R., & Markin, J. S. (1979). Journal of Bacteriology, 137, 51–61.
Becerra, M., Cerdan, E., & Siso, M. I. G. (1998). Biotechnology Techniques, 12, 253–256.
Kim, C. S., Ji, E. S., & Oh, D. K. (2003). Biotechnology Letters, 25, 1769–1774.
Juers, D. H., Matthews, B. W., & Huber, R. E. (2012). Protein Science. doi:10.1002/pro.2165.
Maksimainen, M., Paavilainen, S., Hakulinen, N., & Rouvinen, J. (2012). FEBS Journal, 279, 1788–1798.
Juers, D. H., Heightman, T. D., Vasella, A., McCarter, J. D., Mackenzie, L., Withers, S. G., et al. (2001). Biochemistry, 40, 14781–14794.
Richard, J. P., Huber, R. E., Heo, C., Amyes, T. L., & Lin, S. (1996). Biochemistry, 35, 12387–12401.
Richard, J. P., Huber, R. E., Lin, S., Heo, C., & Amyes, T. L. (1996). Biochemistry, 35, 12377–12386.
Marquardt, D. (1963). SIAM Journal on Applied Mathematics, 11, 431–441.
Cray, J. A., Russell, J. T., Timson, D. J., Singhal, R. S., & Hallsworth, J. E. (2013). Environmental Microbiology. doi:10.1111/1462-2920.12018.
Hallsworth, J. E., Nomura, Y., & Iwahara, I. (1998). Journal of Fermentation and Bioengineering 86, 451–456.
Hallsworth, J. E., Yakimov, M. M., Golyshin, P. N., Gillion, J. L., de D’Auria, G., La Lima Alves, F., et al. (2007). Environmental Microbiology, 9, 801–813.
Sinnott, M. L., & Souchard, I. J. (1973). Biochemical Journal, 133, 89–98.
Richard, J. P., Westerfeld, J. G., Lin, S., & Beard, J. (1995). Biochemistry, 34, 11713–11724.
Cornish-Bowden, A. (2004). Fundamentals of enzyme kinetics. London: Portland Press.
Liu, G. X., Kong, J., Lu, W. W., Kong, W. T., Tian, H., Tian, X. Y., et al. (2011). Journal of Dairy Science, 94, 5811–5820.
Brown, A. D. (1978). Advances in Microbial Physiology, 17, 181–242.
Goodey, N. M., & Benkovic, S. J. (2008). Nature Chemical Biology, 4, 474–482.
Kristiansson, H., & Timson, D. J. (2011). ChemBioChem, 12, 2081–2087.
Brown, A. D. (1990). Microbial water stress physiology. Chichester: Wiley.
Williams, J. P., & Hallsworth, J. E. (2009). Environmental Microbiology, 11, 3292–3308.
Wickson, V. M., & Huber, R. E. (1969). Biochimica et Biophysica Acta, 181, 419–425.
Thomas, K. C., Hynes, S. H., & Ingledew, W. M. (1993). Journal of Industrial Microbiology and Biotechnology, 12, 93–98.
Mansure, J. J., Panek, A. D., Crowe, L. M., & Crowe, J. H. (1994). Biochimica et Biophysica Acta, 1191, 309–316.
Madeira, A., Leitao, L., Soveral, G., Dias, P., Prista, C., Moura, T., et al. (2010). FEMS Yeast Research, 10, 252–258.
Acknowledgments
JEH acknowledges funding from the Biotechnology and Biological Sciences Research Council (project BBF0034711). AWNB received a postgraduate studentship from the Department of Agriculture and Rural Development Northern Ireland and EM received a Nuffield Foundation Science Bursary.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Bell, A.N.W., Magill, E., Hallsworth, J.E. et al. Effects of Alcohols and Compatible Solutes on the Activity of β-Galactosidase. Appl Biochem Biotechnol 169, 786–794 (2013). https://doi.org/10.1007/s12010-012-0003-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-012-0003-3