Purification and Characterization of Trehalase From Acyrthosiphon pisum, a Target for Pest Control

Virgile Neyman, Frédéric Francis, André Matagne, Marc Dieu, Catherine Michaux, Eric Perpete

Research output: Contribution to journalArticlepeer-review


Insect trehalases are glycoside hydrolases essential for trehalose metabolism and stress resistance. We here report the extraction and purification of Acyrthosiphon pisum soluble trehalase (ApTreh-1), its biochemical and structural characterization, as well as the determination of its kinetic properties. The protein has been purified by ammonium sulphate precipitation, first followed by an anion-exchange and then by an affinity chromatography. The SDS-PAGE shows a main band at 70 kDa containing two isoforms of ApTreh-1 (X1 and X2), identified by mass spectrometry and slightly contrasting in the C-terminal region. A phylogenetic tree, a multiple sequence alignment, as well as a modelled 3D-structure were constructed and they all reveal the ApTreh-1 similarity to other insect trehalases, i.e. the two signature motifs 179PGGRFRELYYWDTY192 and 479QWDFPNAWPP489, a glycine-rich region 549GGGGEY554, and the catalytic residues Asp336 and Glu538. The optimum enzyme activity occurs at 45 °C and pH 5.0, with Km and Vmax values of ~ 71 mM and ~ 126 µmol/min/mg, respectively. The present structural and functional characterization of soluble A. pisum trehalase enters the development of new strategies to control the aphids pest without significant risk for non-target organisms and human health.
Original languageEnglish
Pages (from-to)189-200
Number of pages12
JournalThe Protein Journal
Issue number1
Early online date29 Nov 2021
Publication statusPublished - 2022


  • Acyrthosiphon pisum
  • Catalytic and structural properties
  • Molecular modelling
  • Trehalase


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