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Related Experiment Videos

Steady-state fluorescence studies on lipase-vesicle interactions

E W Mosmuller1, E H Pap, A J Visser

  • 1Dept. of Organic Chemistry, Wageningen Agricultural University, The Netherlands.

Biochimica Et Biophysica Acta
|January 3, 1994
PubMed
Summary

Candida cylindracea lipase (CCL) interacts with polymerizable surfactant vesicles. The enzyme incorporates more efficiently into nonpolymerized vesicles, with shallower bilayer penetration in polymerized ones.

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Area of Science:

  • Biochemistry
  • Materials Science
  • Biophysics

Background:

  • Lipases are crucial enzymes in various biological and industrial processes.
  • Surfactant vesicles offer a versatile platform for studying enzyme-membrane interactions.
  • Polymerizable vesicles allow for tunable membrane properties and enhanced stability.

Purpose of the Study:

  • To investigate the interaction of Candida cylindracea lipase (CCL) with positively charged polymerizable surfactant vesicles.
  • To determine the effect of vesicle polymerization on lipase incorporation and membrane penetration.
  • To elucidate the structural changes in vesicles upon lipase interaction.

Main Methods:

  • Steady-state fluorescence techniques were employed.
  • Phase transition temperatures were determined using the membrane probe 1,6-diphenyl-1,3,5-hexatriene (DPH).

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  • Fluorescence anisotropy and resonance energy transfer measured lipase incorporation.
  • Interfacial and internal membrane probes (TMA-DPH and DPH) assessed enzyme penetration depth.

    • Main Results:

    • The phase transition temperature of nonpolymerized vesicles was 49°C, and for polymerized vesicles, it was 45°C.
    • CCL was successfully incorporated into the hydrophobic bilayer of both vesicle types.
    • Lipase incorporation was more efficient in nonpolymerized vesicles compared to polymerized ones.
    • Enzyme penetration into the bilayer was less deep in polymerized vesicles.

    Conclusions:

    • Candida cylindracea lipase interacts with and incorporates into positively charged surfactant vesicles.
    • Vesicle polymerization influences lipase incorporation efficiency and penetration depth.
    • Polymerized vesicles exhibit altered membrane properties affecting enzyme-membrane interactions.