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A multi-pumping flow analysis system for β-galactosidase activity assays.

Justyna Skoczek1, Marta Pokrzywnicka2, Olga Kubacka1

  • 1Faculty of Chemistry, University of Warsaw, L. Pasteura 1, 02-093 Warsaw, Poland.

Food Chemistry
|May 26, 2019
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Summary

A new automated flow analysis system accurately measures beta-galactosidase activity. This system is valuable for analyzing dietary supplements and understanding lactose intolerance.

Keywords:
Dietary supplementsFlow analysisLactose intoleranceβ-Galactosidase

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Enzyme Assays

Background:

  • Beta-galactosidase activity is crucial for lactose metabolism and is relevant in dietary supplements for lactose intolerance.
  • Accurate and efficient determination of enzyme activity is essential for quality control and research.

Purpose of the Study:

  • To develop and validate a fully mechanized Arduino-controlled multi-pumping flow analysis system for determining beta-galactosidase activity.
  • To optimize enzyme detection conditions and evaluate the system's performance for various applications.

Main Methods:

  • A novel bioanalytical method based on photometric detection of p-nitrophenol, a product of enzyme-catalyzed hydrolysis.
  • Utilized an Arduino-controlled multi-pumping system with a flow-through optoelectronic detector.
  • Evaluated optimal pH, temperature, reaction time, substrate selection, and enzyme isoenzyme comparison.

Main Results:

  • The system achieved a wide determination range for beta-galactosidase activity (up to 15 U/mL) with a low detection limit (approx. 0.01 U/mL).
  • Demonstrated high sample throughput (up to 30 detections per hour).
  • Successfully applied to analyze beta-galactosidase activity in dietary supplements and showed potential for amyloglucosidase assays.

Conclusions:

  • The developed automated flow analysis system provides a robust and efficient platform for beta-galactosidase activity determination.
  • The system is suitable for optimizing enzyme assays, comparing enzyme origins, detecting inhibitors, and analyzing enzyme activity in complex matrices like dietary supplements.
  • The system's versatility is further highlighted by its demonstrated utility for amyloglucosidase activity assays.