Comparative study of Bacillus thuringiensis Cry1Ia and Cry1Aa delta-endotoxins: activation process and toxicity against Prays oleae.
Cry1Ia and Cry1Aa proteins exhibited toxicities against Prays oleae with LC(50) of 189 and 116 ng/cm(2), respectively. The ability to process Cry1Ia11 protoxin by trypsin, chymotrypsin and P. oleae larvae proteases was studied and compared to that of Cry1Aa11. After solubilization under high alkaline condition (50mM NaOH), Cry1Aa11 was converted into a major fragment of 65 kDa, whereas Cry1Ia11 protoxin was completely degraded by P. oleae larvae proteases and trypsin and converted into a major fragment of 70 kDa by chymotrypsin. Using less proteases of P. oleae juice, the degradation of Cry1Ia11 was attenuated. When the solubilization (in 50mM Na(2)CO(3) pH 10.5 buffer) and activation were combined, Cry1Ia11 was converted into a proteolytic product of 70 kDa after 3h of incubation with trypsin, chymotrypsin and P. oleae juice. These results suggest that the in vivo solubilization of Cry1Ia11 was assured by larval proteases after a swelling of the corresponding inclusion due to the alkalinity of the larval midgut.
Dammak
Mariam
M
Laboratory of Biopesticides, Centre of Biotechnology of Sfax, Tunisia, P.O. Box 1177, 3061 Sfax, Tunisia.
Tounsi
Slim
S
Rouis
Souad
S
Jaoua
Samir
S
D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism* / Q000633:toxicity*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000378:metabolism* / Q000633:toxicity*; D006460:Hemolysin Proteins / Q000378:metabolism* / Q000633:toxicity*; D009036:Moths / Q000378:metabolism* / Q000469:parasitology*; D011994:Recombinant Proteins / Q000378:metabolism / Q000633:toxicity
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D011994:Recombinant Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis