Activity of alpha-amylase inhibitors from Phaseolus coccineus on digestive alpha-amylases of the coffee berry borer.
Seeds of scarlet runner bean ( Phaseolus coccineus L.) were analyzed for alpha-amylase inhibitor (alpha-AI) activity. Through the use of polyclonal antibodies raised against pure alpha-AI-1 from common bean ( Phaseolus vulgaris L.), typical alpha-AlphaIota polypeptides (Mr 14-18 kDa) as well as a large polypeptide of Mr 32000 Da, usually referred to as "amylase inhibitor like", were detected. The inhibitor activity present in four accessions of P. coccineus was examined, both in semiquantitative zymograms allowing the separation of different isoforms and in quantitative assays against human salivary amylase, porcine pancreatic amylase, and coffee berry borer, Hypothenemus hampei Ferrari (Coleoptera: Scolytidae) amylase. Differential inhibition curves lead to the suggestion that the gene encoding one of the inhibitors in P. coccineus (in accession G35590) would be a good candidate for genetic engineering of coffee resistance toward the coffee berry borer. An in vitro proteolytic digestion treatment of pure alpha-AlphaIota-1 resulted in a rapid loss of the inhibitory activity, seriously affecting its natural capacity to interact with mammalian alpha-amylases.
Valencia-Jiménez
Arnubio
A
Embrapa Recursos Genéticos e Biotecnologia, PqEB, Brasília DF, Brazil. avalbioquimica@yahoo.com
Arboleda Valencia
Jorge W
JW
Grossi-De-Sá
Maria Fátima
MF
D000818:Animals; D001517:Coleoptera / Q000201:enzymology*; D004063:Digestion; D004355:Drug Stability; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D007300:Insect Control / Q000379:methods; D007306:Insecticides; D027805:Phaseolus / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry; D013552:Swine; D000516:alpha-Amylases / Q000037:antagonists & inhibitors*
D004791:Enzyme Inhibitors; D007306:Insecticides; D000516:alpha-Amylases