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Applied and Environmental Microbiology, October 2001, p. 4488-4494, Vol. 67, No. 10
Groupe de recherche en transport membranaire,
Université de Montréal, Montreal, Quebec H3C
3J7,1 and Biotechnology Research
Institute, National Research Council, Montreal, Quebec H4P
2R2,2 Canada
Received 13 February 2001/Accepted 17 July 2001
The effect of pH on the pore-forming ability of two Bacillus
thuringiensis toxins, Cry1Ac and Cry1C, was examined with midgut brush border membrane vesicles isolated from the tobacco hornworm, Manduca sexta, and a light-scattering assay. In the
presence of Cry1Ac, membrane permeability remained high over the entire
pH range tested (6.5 to 10.5) for KCl and tetramethylammonium chloride, but was much lower at pH 6.5 than at higher pHs for potassium gluconate, sucrose, and raffinose. On the other hand, the Cry1C-induced permeability to all substrates tested was much higher at pH 6.5, 7.5, and 8.5 than at pH 9.5 and 10.5. These results indicate that the pores
formed by Cry1Ac are significantly smaller at pH 6.5 than under
alkaline conditions, whereas the pore-forming ability of Cry1C
decreases sharply above pH 8.5. The reduced activity of Cry1C at high
pH correlates well with the fact that its toxicity for M. sexta is considerably weaker than that of Cry1Aa, Cry1Ab, and
Cry1Ac. However, Cry1E, despite having a toxicity comparable to that of
Cry1C, formed channels as efficiently as the Cry1A toxins at pH 10.5. These results strongly suggest that although pH can influence toxin
activity, additional factors also modulate toxin potency in the insect midgut.
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.10.4488-4494.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Differential Effects of pH on the Pore-Forming
Properties of Bacillus thuringiensis Insecticidal
Crystal Toxins
*
Corresponding author. Mailing address: Groupe de
recherche en transport membranaire, Université de Montréal,
P.O. Box 6128, Centre Ville Station, Montreal, Quebec H3C 3J7, Canada.
Phone: (514) 343-7960. Fax: (514) 343-7146. E-mail:
raynald.laprade{at}umontreal.ca.
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