Metabolism of Benzoquinone by Yeast Cells and Oxidative Characteristics of Corresponding Hydroquinone: Application to Highly Sensitive Measurement of Yeast Cell Density by Using Benzoquinone and a Chemiluminescent Probe

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  • TSUKATANI Tadayuki
    Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center
  • IDE Seiji
    Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center
  • UKEDA Hiroyuki
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • MATSUMOTO Kiyoshi
    Institute of Food Biotechnology, Department of Bioscience and Biotechnology, Division of Bioresource and Bioenvironmental Sciences, Graduate School Kyushu University

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The metabolic efficiency of seven derivatives of 1,4-benzoquinone (BQ) by yeast cells and the oxidative characteristics of the corresponding hydroquinones (HQs) were studied by electrochemical, spectrophotometric and chemiluminescent methods. The spectrophotometric method was based on the reduction of a tetrazolium salt to formazan dye during the autoxidation of HQs generated by yeast cells under alkaline conditions. The amounts of HQs detected directly by the electrochemical method did not agree with those calculated from the formazan dye obtained by the spectrophotometric method. A tetrazolium salt was reduced to a formazan dye by both the superoxide anion radical (O2−•) generated during the autoxidation of 2,3,5,6-tetramethyl-1,4-HQ and by HQ itself. Little formazan dye was formed, and hydrogen peroxide (H2O2) was then finally produced during the autoxidation of 1,4-HQ or 2-methyl-1,4-HQ. Formazan dye and H2O2 were generated at a certain ratio during the autoxidation of derivatives of dimethyl-1,4-HQ or 2,3,5-trimethyl-1,4-HQ. The analytical method based on chemiluminescence with lucigenin and 2,3,5,6-tetramethyl-1,4-BQ was applied to highly sensitive measurement of the yeast cell density. A linear relationship between the chemiluminescence intensity and viable cell density was obtained in the range of 1.2×103–4.8×104 cells/ml. The detection limit was 4.8×102 cells/ml.

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