Purification, characterization and antibacterial potential of a lectin isolated from Apuleia leiocarpa seeds

Int J Biol Macromol. 2015 Apr:75:402-8. doi: 10.1016/j.ijbiomac.2015.02.001. Epub 2015 Feb 7.

Abstract

Apuleia leiocarpa is a tree found in Caatinga that has great value in the timber industry. Lectins are carbohydrate-binding proteins with several biotechnological applications. This study shows the isolation, characterization, and antibacterial activity of A. leiocarpa seed lectin (ApulSL). The lectin was chromatographically isolated from a crude extract (in 150 mM NaCl) by using a chitin column. ApulSL adsorbed to the matrix and was eluted using 1.0 M acetic acid. Native ApulSL was characterized as a 55.8-kDa acidic protein. SDS-PAGE showed three polypeptide bands, whereas two-dimensional electrophoresis revealed four spots. The peptides detected by MALDI TOF/TOF did not show sufficient homology (<30%) with the database proteins. Circular dichroism spectroscopy suggested a disordered conformational structure, and fluorescence spectrum showed the presence of tyrosine residues in the hydrophobic core. The hemagglutinating activity of ApulSL was present even after heating to 100 °C, was Mn(2+)-dependent, and inhibited by N-acetylglucosamine, D(-)-arabinose, and azocasein. ApulSL demonstrated bacteriostatic and bactericide effects on gram-positive and gram-negative species, being more effective against three varieties of Xanthomonas campestris (MIC ranging from 11.2 to 22.5 μg/mL and MBC of 22.5 μg/mL). The results of this study reinforce the importance of biochemical prospecting of Caatinga by revealing the antibacterial potential of ApulSL.

Keywords: Apuleia leiocarpa; Lectin; Xanthomonas campestris.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / pharmacology*
  • Bacteria / drug effects
  • Chromatography, Gel
  • Electrophoresis, Gel, Two-Dimensional
  • Fabaceae / chemistry*
  • Hemagglutination / drug effects
  • Hemagglutinins / chemistry
  • Humans
  • Mass Spectrometry
  • Microbial Sensitivity Tests
  • Peptides / chemistry
  • Plant Lectins / chemistry
  • Plant Lectins / isolation & purification*
  • Plant Lectins / pharmacology*
  • Rabbits
  • Seeds / chemistry*

Substances

  • Anti-Bacterial Agents
  • Hemagglutinins
  • Peptides
  • Plant Lectins