The Protein Journal

, Volume 23, Issue 5, pp 343–350 | Cite as

Trypsin Inhibitor from Poecilanthe parviflora Seeds: Purification, Characterization, and Activity Against Pest Proteases

  • Viviane Alves Garcia
  • Maria das Graças Machado Freire
  • José Camillo Novello
  • Sérgio Marangoni
  • Maria Lígia Rodrigues Macedo
Article

Abstract

Plants synthesize a variety of molecules, including proteinaceous proteinase inhibitors, to defend themselves of being attacked by insects. In this work, a novel trypsin inhibitor (PPTI) was purified from the seeds of the native Brazilian tree Poecilanthe parviflora (Benth) (Papilioinodeae, Leguminosae) by gel filtration chromatography on a Sephadex G-100 followed by Superdex G75 chromatography (FPLC), Sepharose 4B-Trypsin column, and fractionated by reversed-phase HPLC on a C-18 column. SDS-PAGE showed that PPTI consisted of a single polypeptide chain with molecular mass of about 16 kDa. The dissociation constant of 1.0 × 10−7 M was obtained with bovine trypsin. PPTI was stable over a wide range of temperature and pH and in the presence of DTT. The N-terminal sequence of the PPTI showed a high degree of homology with other Kunitz-type inhibitors. Trypsin-like activity in midguts of larval Diatraea saccharalis, Anagasta kuehniella, Spodoptera frugiperda, and Corcyra cephalonica were substantially inhibited by PPTI.

Kunitz inhibitor Leguminosae pest protease Poecilanthe parviflora trypsin inhibitor 

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Copyright information

© Plenum Publishing Corporation 2004

Authors and Affiliations

  • Viviane Alves Garcia
  • Maria das Graças Machado Freire
  • José Camillo Novello
  • Sérgio Marangoni
  • Maria Lígia Rodrigues Macedo

There are no affiliations available

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