Effects of Alcohols and Compatible Solutes on the Activity of β-Galactosidase

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• 作者：Andrew N. W. Bell (1)
  Emma Magill (1)
  John E. Hallsworth (1)
  David J. Timson (1)
  
• 关键词：Protein denaturation ; Chaotropic solute ; Model enzyme ; Steady ; state kinetics ; Polyol ; Enzyme flexibility
• 刊名：Applied Biochemistry and Biotechnology
• 出版年：2013
• 出版时间：February 2013
• 年：2013
• 卷：169
• 期：3
• 页码：786-794
• 全文大小：336KB
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• 作者单位：Andrew N. W. Bell (1)
  Emma Magill (1)
  John E. Hallsworth (1)
  David J. Timson (1)
  
  1. School of Biological Sciences, Medical Biology Centre, Queen’s University Belfast, 97 Lisburn Road, Belfast, BT9 7BL, UK
  
• ISSN：1559-0291

文摘

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.