A trypsin inhibitor from Sapindus saponaria L. seeds: purification, characterization, and activity towards pest insect digestive enzyme

Protein J. 2011 Jan;30(1):9-19. doi: 10.1007/s10930-010-9296-7.

Abstract

The present paper describes the purification, characterization and determination of the partial primary structure of the first trypsin inhibitor isolated from the family Sapindaceae. A highly stable, potent trypsin inhibitor (SSTI) was purified to homogeneity. SDS-PAGE analysis revealed that the protein consists of a two-polypeptide chain with molecular masses of approximately 15 and 3 kDa. The purified inhibitor inhibited bovine trypsin at a 1:1 M ratio. Kinetic analysis revealed that the protein is a competitive inhibitor with an equilibrium dissociation constant of 10⁻⁹ M for trypsin. The partial NH₂- terminal sequence of 36 amino acids in SSTI indicates homology with other members of the trypsin-inhibitor family from different sources. This inhibitor is highly stable in the presence of denaturing agents. SSTI showed significant inhibitory activity against trypsin-like proteases present in the larval midgut on Anagasta kuehniella, Corcyra cephalonica, Diatreae saccharalis and Anticarsia gemmatalis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arginine / chemistry
  • Brazil
  • Dithiothreitol / chemistry
  • Dithiothreitol / pharmacology
  • Drug Stability
  • Hydrogen-Ion Concentration
  • Insecticides / chemistry
  • Insecticides / isolation & purification*
  • Insecticides / pharmacology
  • Intestine, Small / enzymology
  • Kinetics
  • Larva / enzymology
  • Lepidoptera*
  • Molecular Sequence Data
  • Molecular Weight
  • Plant Proteins / chemistry
  • Plant Proteins / isolation & purification*
  • Plant Proteins / pharmacology
  • Protein Stability
  • Sapindus / chemistry*
  • Seeds / chemistry*
  • Trypsin Inhibitors / chemistry
  • Trypsin Inhibitors / isolation & purification*
  • Trypsin Inhibitors / pharmacology

Substances

  • Insecticides
  • Plant Proteins
  • Trypsin Inhibitors
  • Arginine
  • Dithiothreitol