Time-resolved fluorescence observation of di-tyrosine formation in horseradish peroxidase upon ultrasound treatment leading to enzyme inactivation

Spectrochim Acta A Mol Biomol Spectrosc. 2017 Feb 15:173:324-327. doi: 10.1016/j.saa.2016.09.035. Epub 2016 Sep 20.

Abstract

The application of ultrasound to a solution can induce cavitional phenomena and generate high localised temperatures and pressures. These are dependent of the frequency used and have enabled ultrasound application in areas such as synthetic, green and food chemistry. High frequency (100kHz to 1MHz) in particular is promising in food chemistry as a means to inactivate enzymes, replacing the need to use periods of high temperature. A plant enzyme, horseradish peroxidase, was studied using time-resolved fluorescence techniques as a means to assess the effect of high frequency (378kHz and 583kHz) ultrasound treatment at equivalent acoustic powers. This uncovered the fluorescence emission from a newly formed species, attributed to the formation of di-tyrosine within the horseradish peroxidase structure caused by auto-oxidation, and linked to enzyme inactivation.

Keywords: Decay associated spectra; Excitation-emission matrix; High frequency ultrasound; Metal catalysed oxidation; Time-resolved emission spectrum.

MeSH terms

  • Enzyme Activation
  • Horseradish Peroxidase / chemistry*
  • Horseradish Peroxidase / metabolism
  • Protein Denaturation
  • Solvents / chemistry
  • Sonication
  • Spectrometry, Fluorescence / methods*
  • Tyrosine / chemistry*
  • Ultrasonics / methods*

Substances

  • Solvents
  • Tyrosine
  • Horseradish Peroxidase