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Related Experiment Videos

Second harmonic generation of glucose oxidase at the air/water interface.

J Rinuy1, P F Brevet, H H Girault

  • 1Laboratoire d'Electrochimie, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.

Biophysical Journal
|December 10, 1999
PubMed
Summary
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Glucose oxidase (GOx) enzyme adsorption at the air/water interface was studied using surface second harmonic generation (SSHG). Researchers observed GOx reorientation at high concentrations and its interaction with charged surfactants.

Area of Science:

  • Biophysics
  • Surface Science
  • Enzyme Kinetics

Background:

  • Enzyme adsorption at interfaces is crucial for biosensor development.
  • Understanding protein behavior at the air/water interface informs biomaterial design.
  • Glucose oxidase (GOx) is a key enzyme in biotechnological applications.

Purpose of the Study:

  • To investigate the adsorption behavior of glucose oxidase (GOx) at the air/water interface.
  • To monitor GOx surface concentration and orientation using nonlinear optics.
  • To explore the influence of charged surfactants on GOx adsorption.

Main Methods:

  • Nonlinear optical technique: surface second harmonic generation (SSHG).
  • Resonant SSHG probing the flavin adenine dinucleotide (FAD) chromophore in GOx.

Related Experiment Videos

  • Monitoring protein adsorption and orientation changes at varying concentrations and surfactant conditions.

    • Main Results:

    • GOx adsorption was detectable at bulk concentrations as low as 70 nM due to resonance enhancement.
    • A change in FAD chromophore orientation was observed at high GOx coverage, indicating protein rearrangement or reorientation.
    • Positively charged surfactants increased GOx surface concentration, while negatively charged surfactants decreased it, returning the signal to baseline.

    Conclusions:

    • SSHG is a sensitive technique for studying enzyme adsorption and conformational changes at interfaces.
    • GOx exhibits concentration-dependent structural changes at the air/water interface.
    • Charged surfactants modulate GOx adsorption, offering potential for interface control in biotechnological systems.