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Author (up) Silva, D.P.; Casado-Filho, E.L.; Correa, A.S.R.; Farias, L.R.; Bloch, C.J.; de Sa, M.F.G.; Mendes, P.A.M.; Quirino, B.F.; Noronha, E.F.; Franco, O.L. url  doi
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  Title Identification of an alpha-amylase inhibitor from Pterodon pubescens with ability to inhibit cowpea weevil digestive enzymes Type Journal Article
  Year 2007 Publication Journal of Agricultural and Food Chemistry Abbreviated Journal J Agric Food Chem  
  Volume 55 Issue 11 Pages 4382-4387  
  Keywords Animals; Biological Assay; Enzyme Inhibitors/chemistry/*isolation & purification/*pharmacology; Fabaceae/*chemistry; Female; Male; Pest Control, Biological/methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Weevils/*enzymology; alpha-Amylases/*antagonists & inhibitors  
  Abstract Cowpea seeds (Vigna ungiculata) are widely cultivated by poor farmers in Latin America and Africa and are often severely damaged by the cowpea weevil Callosobruchus maculatus. A proteinaceous inhibitor of cowpea weevil digestive enzymes, PpAI, was purified from white sucupira seeds (Pterodon pubescens) and biochemically characterized in this study. Proteins were extracted from seeds and precipitated with ammonium sulfate at 100% saturation. This fraction was applied onto a Red-sepharose CL-6B column, and the retained peak showed 70% inhibitory activity toward larval C. maculatus digestive alpha-amylases. The retained peak was then purified using an analytical reversed-phase HPLC column. Purified PpAI showed 65% inhibitory activity against larval C. maculatus enzymes. Enzymatic assays also showed that the purified P. pubescens inhibitor was unable to reduce the activity of mammalian alpha-amylases, suggesting specificity toward insect enzymes. Moreover, artificial seeds containing PpAI were able to reduce larval weight by 36% and cause 55% mortality. Mass spectrometry and SDS-PAGE analyses indicated that PpAI showed a molecular mass of approximately 5.0 kDa. This alpha-amylase inhibitor, coming from a native Cerrado plant, could be used to construct a genetically engineered cowpea with enhanced resistance against weevil pests.  
  Address Centro de AnAlises Proteomicas e Bioquimicas, Programa de Pos-Graduacao em Ciencias Genomicas e Biotecnologia, Brasilia DF, Brazil  
  Corporate Author Thesis  
  Publisher Place of Publication Editor  
  Language English Summary Language Original Title  
  Series Editor Series Title Abbreviated Series Title  
  Series Volume Series Issue Edition  
  ISSN 0021-8561 ISBN Medium  
  Area Expedition Conference  
  Notes PMID:17488029 Approved no  
  Call Number GGC @ adcosta @ Serial 35  
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