Kinetic and Mechanistic Studies of Oxidation of Vitamin a Alcohol to Vitamin a Aldehyde by Horse Liver Alcohol Dehydrogenase. The Inhibition by Ethanol and Pyrazole

  • Y. Pocker
  • K. W. Raymond


The present investigation shows that all-trans retinol (Vitamin A alcohol), an alcohol of great physiological importance, is efficiently oxidized to all-trans retinaldehyde by the enzyme horse liver alcohol dehydrogenase. We observe a Km retinol value of 145μM and a turnover number of 0.45s−1 for the oxidation of all-trans retinol in the presence of Triton X-100, a surfactant used as a solubilizer. Over the concentration range of surfactant used (up to 0.1% Triton X-100) our studies on the oxidation of ethanol and all-trans retinol show that turnover numbers for both reactions remain constant as does the value for Km ethanol. On increasing the concentration of Triton X-100 from 0.025% to 0.10%, however, the Km retinol value increases by a factor of two. This behavior for retinol oxidation can be attributed to the partitioning of retinol between enzyme and surfactant. Pyrazole, a known inhibitor of alcohol oxidation by horse liver alcohol dehydrogenase, is a competitive inhibitor of both all-trans retinol and ethanol, with observed Ki values of 3.3×10−7 M and 3.9×10−7 M, respectively. We also find that ethanol inhibits all-trans retinol oxidation in a complex fashion, an observation which may have important consequences in view of the physiological role of retinol and its oxidation products. Our present studies indicate that all-trans retinol binds in the same region of the enzyme as does ethanol and is oxidized with an efficiency approaching that of ethanol itself.


Ethanol Concentration Surfactant Concentration Alcohol Dehydrogenase Turnover Number Ethanol Oxidation 
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Copyright information

© Springer Science+Business Media New York 1980

Authors and Affiliations

  • Y. Pocker
    • 1
  • K. W. Raymond
    • 1
  1. 1.Department of ChemistryUniversity of WashingtonSeattleUSA

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