<b>Purification of a membrane-bound trypsin-like enzyme from the gut of the velvetbean caterpillar (<i>Anticarsia gemmatalis </i> Hübner)</b> - doi: 10.4025/actascibiolsci.v34i3.9239

  • Denise Torres Cruz Reis Universidade Federal de Viçosa
  • Thiago Rennó dos Mares-Guia Universidade de São Paulo
  • Jamil Silvano de Oliveira Universidade Federal de Minas Gerais
  • Alexandre Martins Costa Santos Universidade Federal de Minas Gerais
  • Marcelo Matos Santoro Universidade Federal de Minas Gerais
  • Maria Goreti Almeida Oliveira Universidade Federal de Viçosa
Palavras-chave: insect-plant interaction, soybean pest control, trypsin-like enzyme purification

Resumo

Disruption of protein digestion in insects by specific endoprotease inhibitors is being regarded as an alternative to conventional insecticides for pest control. To optimize the effectiveness of this strategy, the understanding of the endoprotease diversity of the target insect is crucial. In this sense, a membrane-bound trypsin-like enzyme from the gut of Anticarsia gemmatalis fifth-instar larvae was purified. Non-soluble fraction of the gut extract was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) and subjected to a p-aminobenzamidine affinity chromatography followed by anion-exchange chromatography. The yield of the purified enzyme was 11% with a purification factor of 143 and a final specific activity of 18.6 µM min.-1 mg-1 protein using N-α-benzoyl-L- Arg-p-nitroanilide (L-BApNA) as substrate. The purified sample showed a single band with proteolytic activity active and apparent molecular mass of 25 kDa on SDS-PAGE. Molecular mass determined by MALDI-TOF mass spectrometry was 28,632 ± 26 Da. Although the low recovery and the difficulties in purifying large enzyme amounts limited its further characterization, the results contribute for the understanding of the proteases present on A. gemmatalis gut, which are potential targets for natural or specifically designed protease inhibitors.

Downloads

Não há dados estatísticos.

Biografia do Autor

Denise Torres Cruz Reis, Universidade Federal de Viçosa
Departamento de Bioquímica e Biologia Molecular
Publicado
2012-02-14
Como Citar
Reis, D. T. C., Mares-Guia, T. R. dos, Oliveira, J. S. de, Santos, A. M. C., Santoro, M. M., & Oliveira, M. G. A. (2012). <b>Purification of a membrane-bound trypsin-like enzyme from the gut of the velvetbean caterpillar (<i>Anticarsia gemmatalis </i> Hübner)</b&gt; - doi: 10.4025/actascibiolsci.v34i3.9239. Acta Scientiarum. Biological Sciences, 34(3), 263-270. https://doi.org/10.4025/actascibiolsci.v34i3.9239
Seção
Biotecnologia

 

0.6
2019CiteScore
 
 
31st percentile
Powered by  Scopus

 

 

0.6
2019CiteScore
 
 
31st percentile
Powered by  Scopus