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EVALUATION OF IN VITRO AND IN VIVO EFFECTS OF SEMIPURIFIED PROTEINASE INHIBITORS FROM T HEOBROMA SEEDS ON MIDGUT PROTEASE ACTIVITY OF L EPIDOPTERAN PEST INSECTS
Author(s) -
Paulillo Luis Cesar Maffei Sartini,
Magno Sebbenn Alexandre,
de Carvalho Derbyshire Maria Tereza Vitral,
GóesNeto Aristóteles,
de Paula Brotto Marco Aurélio,
Figueira Antonio
Publication year - 2012
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.21038
Subject(s) - trypsin , chymotrypsin , biology , midgut , protease , biochemistry , kunitz sti protease inhibitor , in vitro , in vivo , enzyme , spodoptera , microbiology and biotechnology , botany , recombinant dna , larva , gene
We have characterized in vitro and in vivo effects of trypsin inhibitors from T heobroma seeds on the activity of trypsin‐ and chymotrypsin‐like proteins from L epidopteran pest insects. The action of semipurified trypsin inhibitors from T heobroma 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 T heobroma 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 T heobroma seeds significantly inhibited purified bovine trypsin. With respect to the influence of T heobroma trypsin inhibitors on intact insects, the inclusion of T . cacao extracts in artificial diets of velvet bean caterpillars ( A nticarsia gemmatalis) and sugarcane borer ( D iatraea 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 .