Construction and Characterization of Glucose Enzyme Sensor Employing Engineered Water Soluble PQQ Glucose Dehydrogenase with Improved Thermal Stability

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  • TAKAHASHI Yuka
    Department of Biotechnology, Tokyo University of Agriculture and Technology
  • IGARASHI Satoshi
    Department of Biotechnology, Tokyo University of Agriculture and Technology
  • NAKAZAWA Yukie
    Department of Biotechnology, Tokyo University of Agriculture and Technology
  • TSUGAWA Wakako
    Department of Biotechnology, Tokyo University of Agriculture and Technology
  • SODE Koji
    Department of Biotechnology, Tokyo University of Agriculture and Technology

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Abstract

<p>In this study, we constructed glucose enzyme sensors with increased thermal stability employing engineered PQQGDH-B. First, we demonstrated the enzyme glucose sensor employing an engineered PQQGDH-B, Ser231 Lys. The residual activity after heat treatment at 60°C for 2 hours of the enzyme electrode employed Ser231Lys was 80% of the initial activity, whereas the electrode employed native PQQGDH-B was 30%. Second, we investigated the effect of cross-linking chemical modification on the thermal stability of PQQGDH-B. The cross-linked PQQGDH-B had high thermal stability and the half life time at 55°C was 63 min whereas that of native enzyme was 4 min. Furthermore, we demonstrated the Flow Injection Analysis (FIA) system for glucose measurement employed the cross-linked PQQGDH-B. The high operational stability of this system showed the very stable responses (SD was within 3%) for 100 times of glucose injection.</p>

Journal

  • Electrochemistry

    Electrochemistry 68 (11), 907-911, 2000-11-05

    The Electrochemical Society of Japan

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