Trypsin-like activity of membrane-bound midgut proteases from Anticarsia gemmatalis (Lepidoptera: Noctuidae)
| Autor: | Luciana Pereira XAVIER, Maria Goreti ALMEIDA OLIVEIRA, Raul Narciso Carvalho GUEDES, Agenor Valarades SANTOS, Salvatore Giovanni DE SIMONE |
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| Jazyk: | angličtina |
| Rok vydání: | 2005 |
| Předmět: | |
| Zdroj: | European Journal of Entomology, Vol 102, Iss 2, Pp 147-153 (2005) |
| Druh dokumentu: | article |
| ISSN: | 1210-5759 1802-8829 |
| DOI: | 10.14411/eje.2005.023 |
| Popis: | Membrane-bound proteases from preparations of the midgut of 5th instar velvetbean caterpillars, Anticarsia gemmatalis (Hübner) were obtained by resuspension of the pellet obtained after 100,000 g centrifugation. As expected of trypsin-like proteases, they hydrolyzed casein and the synthetic substrates N-α-benzoyl-L-Arg-p-nitroanilidine (L-BApNA) and N-α-p-tosyl-L-Arg methyl ester (L-TAME). Higher activities were observed at 50°C, and at pH 8.5 and 8.0 for both synthetic substrates L-BApNA and L-TAME. The membrane-bound proteases were inhibited by EDTA, phenylmethan sulphonyl fluoride (PMSF), tosyl-L-lysine chloromethyl ketone (TLCK), benzamidine and aprotinin. TLCK and benzamidine were particularly active inhibitors. The KM-values obtained were 0.23 mM for L-BApNA and 92.5 µM for L-TAME. These results provide evidence for the presence of membrane-bound trypsin-like proteases in the midgut of the velvetbean caterpillar, a key soybean pest in warm climates. The interaction between A. gemmatalis digestive proteases and soybean protease inhibitors has potentially important consequences for soybean breeding programs. |
| Databáze: | Directory of Open Access Journals |
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