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Related Experiment Videos

Extended-range glucose sensor employing engineered glucose dehydrogenases.

T Yamazaki1, K Kojima, K Sode

  • 1Department of Biotechnology, Tokyo University of Agriculture and Technology, Nakamachi, Koganei, Japan.

Analytical Chemistry
|October 12, 2000
PubMed
Summary

This study introduces a novel enzyme glucose sensor strategy using protein-engineered enzymes to expand the dynamic range. The developed sensor accurately measures glucose concentrations over a significantly wider range than conventional methods.

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Area of Science:

  • Biotechnology
  • Enzyme Engineering
  • Biosensors

Background:

  • Traditional glucose sensors have limited dynamic ranges, hindering accurate measurements across diverse concentrations.
  • Protein engineering offers a pathway to modify enzyme kinetics, such as the Michaelis constant (Km), for improved sensor performance.

Purpose of the Study:

  • To develop an enzyme glucose sensor with an expanded dynamic range using protein-engineered enzymes.
  • To evaluate the performance of a composite colorimetric and amperometric glucose sensor system.

Main Methods:

  • Engineered Escherichia coli pyrroloquinoline quinone glucose dehydrogenase (PQQGDH) with a His775Asp substitution to increase Km.
  • Constructed composite colorimetric and amperometric analytical systems using wild-type and engineered PQQGDH variants.

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  • Evaluated the dynamic range, accuracy, and substrate specificity of the developed sensor systems.

    • Main Results:

    • The composite colorimetric sensor demonstrated a wide dynamic range of 0.5–30 mM with <5% error.
    • The composite amperometric sensor achieved an expanded dynamic range of 3–70 mM, encompassing single-enzyme ranges.
    • The engineered enzymes provided narrower substrate specificity for glucose detection.

    Conclusions:

    • Protein engineering of PQQGDH with altered Michaelis constants is effective for expanding glucose sensor dynamic range.
    • Composite enzyme systems offer enhanced glucose measurement capabilities, overcoming limitations of single-enzyme sensors.
    • The developed extended-range glucose sensors show promise for accurate and versatile glucose monitoring.