Probing thermal behaviour of microbial transglutaminase with fluorescence and in silico methods

J Sci Food Agric. 2013 Mar 15;93(4):794-802. doi: 10.1002/jsfa.5799. Epub 2012 Jul 27.

Abstract

Background: Knowledge of transglutaminase behaviour at thermal treatment allows efficient applications in food processing. The heat-induced conformational changes of microbial transglutaminase were studied by fluorescence spectroscopy and a molecular modelling approach.

Results: The experimental results indicate the unfolding of transglutaminase in a single-phase reaction, at temperatures over 60 °C. The incidence of conformational changes is also supported by the increase of both intrinsic and 1-anilino-8-naphthalene sulfonate fluorescence intensity with temperature. Changes in the secondary and tertiary structure of transglutaminase were outlined after running molecular dynamics simulations at temperatures ranging from 25 °C to 80 °C.

Conclusion: The motif's particularities varied with the temperature, suggesting structural rearrangements of the protein, mainly in helices. The largest deviation from the structure equilibrated at 25 °C was observed at 80 °C.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Computer Simulation
  • Food Handling
  • Food Microbiology*
  • Hot Temperature*
  • Humans
  • Protein Denaturation
  • Protein Folding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Spectrometry, Fluorescence
  • Transglutaminases / chemistry*

Substances

  • Bacterial Proteins
  • Transglutaminases