EVALUATION OF IN VITRO AND IN VIVO EFFECTS OF SEMIPURIFIED PROTEINASE INHIBITORS FROM THEOBROMA SEEDS ON MIDGUT PROTEASE ACTIVITY OF LEPIDOPTERAN PEST INSECTS

Authors

  • Luis Cesar Maffei Sartini Paulillo,

    Corresponding author
    • Faculdade de Tecnologia e Ciências, Laboratório de Biotecnologia e Microbiologia, Salvador, Bahia, Brazil
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  • Alexandre Magno Sebbenn,

    1. Instituto Florestal do Estado de São Paulo, Estação Experimental de Tupi, Piracicaba, São Paulo, Brazil
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  • Maria Tereza Vitral de Carvalho Derbyshire,

    1. Laboratório de Melhoramento de Plantas, Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Piracicaba, São Paulo, Brazil
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  • Aristóteles Góes-Neto,

    1. Laboratório de Pesquisa em Microbiologia, Departamento de Ciências Biológicas, Universidade Estadual de Feira de Santana, Feira de Santana, Bahia, Brazil
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  • Marco Aurélio de Paula Brotto,

    1. Muscle Biology Research Group-MUBIG, University of Missouri-Kansas City, Kansas City, Missouri
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  • Antonio Figueira

    1. Laboratório de Melhoramento de Plantas, Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Piracicaba, São Paulo, Brazil
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  • Grant sponsor: Foundation for Research Support of the State of São Paulo (FAPESP); Grant sponsor: Universidade Estadual de Feira de Santana (UEFS); Grant sponsor: Conselho Nacional de Pesquisa-Brazil (CNPq); Grant number: 314505/2009-6; Grant sponsor: NIH-NIAMS; Grant number: 1RC2AR058963-01; Grant sponsor: Missouri Life Sciences Research Board.

Correspondence to: Luis Cesar M. S. Paulillo, Faculdade de Tecnologia e Ciências, Mestrado Profissional em Bioenergia, Laboratório de Biotecnologia e Microbiologia, 41820-785, Salvador, Bahia, Brazil.

E-mail: lpaulillo.ssa@ftc.br or lcpaulillo@gmail.com

Abstract

We have characterized in vitro and in vivo effects of trypsin inhibitors from Theobroma seeds on the activity of trypsin- and chymotrypsin-like proteins from Lepidopteran pest insects. The action of semipurified trypsin inhibitors from Theobroma was evaluated by the inhibition of bovine trypsin and chymotrypsin activities determined by the hydrolysis of N-Benzoyl-DL-Arginine-p-Nitroanilide (BAPA) and N-Succinyl-Ala-Ala-Pho-Phe p-Nitroanilide (S-(Ala)2ProPhe-pNA). Proteinase inhibitor activities from Theobroma cacao and T. obovatum seeds were the most effective in inhibiting trypsin-like proteins, whereas those from T. obovatum and T. sylvestre were the most efficient against chymotrypsin-like proteins. All larvae midgut extracts showed trypsin-like proteolytic activities, and the putative trypsin inhibitors from Theobroma seeds significantly inhibited purified bovine trypsin. With respect to the influence of Theobroma trypsin inhibitors on intact insects, the inclusion of T. cacao extracts in artificial diets of velvet bean caterpillars (Anticarsia gemmatalis) and sugarcane borer (Diatraea saccharalis) produced a significant increase in the percentage of adult deformation, which is directly related to both the survival rate of the insects and oviposition.

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