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A simple and sensitive method for the estimation of microbial lipase activity.

K Veeraragavan1

  • 1Biochemical Engineering Section, National Research Council of Canada, Montreal, Quebec.

Analytical Biochemistry
|May 1, 1990
PubMed
Summary

This study introduces a simple and sensitive high-performance liquid chromatography method for estimating microbial lipase activity. Gum arabic proved optimal for substrate preparation, offering a superior alternative to titration and reversed micelles methods.

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Area of Science:

  • Biochemistry
  • Enzymology
  • Analytical Chemistry

Background:

  • Microbial lipases are crucial enzymes with diverse industrial applications.
  • Accurate and sensitive quantification of lipase activity is essential for enzyme characterization and process optimization.
  • Existing methods for lipase activity estimation have limitations in sensitivity and simplicity.

Purpose of the Study:

  • To develop and validate a simple, sensitive, and reliable method for quantifying microbial lipase activity.
  • To optimize the preparation of emulsified substrates for lipase assays.
  • To compare the performance of the developed method with established techniques.

Main Methods:

  • Lipase was incubated with an emulsified triolein substrate prepared using various solubilizers (e.g., gum arabic, Triton X-100).

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  • Fatty acid products, specifically oleic acid, were quantified using high-performance liquid chromatography (HPLC) on an ODS column.
  • The effects of mono- and divalent ions on enzyme activity were analyzed.

    • Main Results:

    • Gum arabic was identified as the most effective solubilizer for preparing the emulsified substrate.
    • The developed HPLC method demonstrated high sensitivity and simplicity compared to titration and reversed micelles methods.
    • The influence of various ions on microbial lipase activity was elucidated.

    Conclusions:

    • The novel HPLC-based method offers a significant advancement for accurate microbial lipase activity determination.
    • Optimized substrate preparation using gum arabic enhances assay reliability.
    • This method provides a valuable tool for researchers and industries working with microbial lipases.