PROTEOLYTIC ACTIVITY OF AFRICANIZED HONEYBEE (Apis mellifera: HYMENOPTERA, APIDAE) VENOM

P. R. M. DE LIMA(1), M. R., BROCHETTO-BRAGA(1,2,3), J. CHAUD-NETTO(1,2,3)

(1)Department of Biology, Institute of Biosciences of Rio Claro, State of São Paulo, Brazil; (2)Center of Study of Social Insects - CEIS - UNESP, State of São Paulo, Brazil; (3)Center for the Study of Venoms and Venomous Animals - CEVAP-UNESP, State of São Paulo, Brazil.

ABSTRACT: Some properties of a Africanized honeybee venom proteases were determined by enzymatic assays in solution, electrophoresis in SDS-PAGE, and gel filtration. Bee venom extracts were obtained by reservoir disruption, selective dialysis (cut off 12 kDa) to eliminate small components, such as the protease inhibitor present in the venom, and then fractionation of the dialyzed extract by gel filtration on a Sephadex G-100 column. The optimal conditions for the caseinolytic assays were pH 9.5, 2-hour digestion at 37°C, and 1% casein concentration. The proteolytic activity was also determined by electrophoresis in SDS-PAGE with co-polymerized gelatin with three major bands of 66.0, 41.6, and 25.1 kDa. A principal serine-protease-like mechanism was revealed in the enriched fraction of proteolytic activity.

KEY WORDS: venom, africanized bee, Apis mellifera, proteolytic activity, gel filtration chromatography.

CORRESPONDENCE TO:

M. R. BROCHETTO-BRAGA - Departamento de Biologia do Instituto de Biociências, Caixa Postal 199, CEP 13506-900, Rio Claro, São Paulo, Brazil. Email mrbbraga@rc.unesp.br