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Study on multiple-enzyme electrode for sucrose determination.

W Hu1, X Zhang, X Zhang

  • 1Wuhan Institute of Virology, Chinese Academy of Sciences, Hubei.

Chinese Journal of Biotechnology
|January 1, 1991
PubMed
Summary
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A novel enzyme electrode coimmobilizing invertase (INV), mutarotase (MUT), and glucose oxidase (GOD) was developed for accurate sucrose detection. This biosensor demonstrates enhanced stability and rapid response for analyzing sucrose in fermentation broths.

Area of Science:

  • Biotechnology
  • Biosensor Development
  • Enzyme Engineering

Background:

  • Accurate and rapid determination of sucrose concentration is crucial for industrial processes, particularly in fermentation.
  • Existing biosensors often face challenges with stability, response time, and interference from related compounds like glucose.

Purpose of the Study:

  • To develop and characterize a novel multi-enzyme electrode for sensitive and selective sucrose detection.
  • To optimize enzyme composition and operational parameters for enhanced sensor performance.

Main Methods:

  • Coimmobilization of invertase (INV), mutarotase (MUT), and glucose oxidase (GOD) onto a teflon membrane using glutaraldehyde-bridged covalent bonding.
  • Integration of the enzyme membrane with an oxygen electrode in a flow injection analysis system (EFIA).

Related Experiment Videos

  • Optimization of enzyme ratios, pH, and temperature for maximal sensor activity and stability.

    • Main Results:

    • Optimal enzyme ratio (INV:MUT:GOD) determined as 72:48:2.4, achieving INV-MUT recovery >42.9%.
    • Optimal sensor activity observed at pH 5.8-6.5 and temperature 35-45°C.
    • Demonstrated excellent linearity for sucrose detection (5x10⁻⁴–10⁻¹ M kinetic, 10⁻⁵–2x10⁻³ M steady state) with rapid response times (20s kinetic, 2 min steady state).
    • Achieved high accuracy (98% recovery) in fermentation broth analysis, with interference from glucose effectively eliminated.
    • Exhibited remarkable operational stability (>120 hours) and storage stability (30 days).

    Conclusions:

    • The developed multi-enzyme electrode offers a stable, sensitive, and rapid platform for sucrose quantification.
    • The improved lifetime and stability surpass previous reports, making it suitable for industrial applications.
    • This biosensor provides a reliable method for monitoring sucrose in complex matrices like fermentation broths.