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Integrity of the 130 kda polypeptide of Bacillus Thuringiensis SSP. Israelensis δ-Endotoxin in K-S sporulation medium

Published online by Cambridge University Press:  19 September 2011

Z. Ngalo Otieno-Ayayo
Affiliation:
Department of Life Sciences, Ben Gurion University of the Negev P.O. Box 653, Beer Sheva 84105, Israel
David M. Chipman
Affiliation:
Department of Life Sciences, Ben Gurion University of the Negev P.O. Box 653, Beer Sheva 84105, Israel
Arieh Zaritsky
Affiliation:
Department of Life Sciences, Ben Gurion University of the Negev P.O. Box 653, Beer Sheva 84105, Israel
Kamal Khawaled
Affiliation:
Department of Life Sciences, Ben Gurion University of the Negev P.O. Box 653, Beer Sheva 84105, Israel
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Abstract

The 130 kDa polypeptide appears to be the most susceptible of the polypeptides constituting the Bacillus thuringiensis ssp. israelensis δ-endotoxin to the proteases associated with sporulation. Stabilization of this polypeptide in the recently described K-S sporulation medium has been achieved through limiting its cleavage by removing contaminating proteases from the culture by washing. In this way, the complete δ-endotoxin profile is retained for studies on all of the constituent polypeptide fractions.

Résumé

Parmi les polypeptides constituant δ-endotoxine de Bacillus thuringiensis israelensis, le polypeptide de poids moléculaire 130 kDa semble étre le plus vulnérable aux proteases associées à la sporulation. La stabilisation de ce polypeptide par le procéde récemment décrit, “milieu de sporulation K-S”, a été obtenue en limitant sa dégradation par lavage des protéase ayant contaminées de milieu de culture. Ainsi, le profile complet de δ-endotoxine a été retenu par études sur toutes les composantes des polypeptides.

Type
Research Artilces
Copyright
Copyright © ICIPE 1993

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References

REFERENCES

Ang, B. J. and Nickerson, K. W. (1978) Purification of the protein crystal from Bacillus thuringiensis by Izonal gradient centrifugation. Appi Environ. Microbiol. 36, 625626.CrossRefGoogle ScholarPubMed
Bodanszky, M. and Martinez, J. (1983) Side Reactions in Peptide Synthesis, in The Peptides: Analysis, Synthesis, Biology (Edited by Gross, Erhard and Meinnofer, Johannes), Vol. 5 pp. 111216. Academic Press Inc.Google Scholar
Bulla, L. A. Jr, B'echtel, D. B., Kramer, K. J., Sethna, Y. I., Aronson, A. I. and Fitz-James, P. C. (1980) Ultrastructure, physiology and biochemistry of Bacillus thuringiensis. CRC. Crit. Rev. Microbiol. 8, 147204.CrossRefGoogle ScholarPubMed
Chilcott, C. N., Kalmakoff, J. and Pillai, J. S. (1983) Characterization of proteolytic activity associated with Bacillus thuringiensis var israelensis crystals. FEMS Microbiol. Lett. 18, 3741.CrossRefGoogle Scholar
Goodman, N. S., Gottfried, R. J. and Rogoff, M. H. (1967) Biphasic system of separation of spores and crystals of Bacillus thuringiensis. J. Bacteriol. 94, 485.CrossRefGoogle ScholarPubMed
Hanson, R. S., Peterson, J. A., and Yousten, A. A. (1970) Unique biochemical events in bacterial sporulation. Annu. Rev. Microbiol. 24, 5390.CrossRefGoogle ScholarPubMed
Manillon, J. and Delcour, J. (1984) A convenient procedure for the preparation of highly purified parasporal crystals of Bacillus thuringiensis. J. Microbiol. Methods 3, 6975.CrossRefGoogle Scholar
Nickerson, K. W. and Swanson, J. D. ( 1981 ) Removal of contaminating proteases from Bacillus thuringiensis parasporal crystals by density gradient centrifugation in NaBr. European J. Appi. Microbiol. Biotechnol. 13, 213215.CrossRefGoogle Scholar
Otieno-Ayayo, Z. N., Khawaled, K., Zaritsky, A. and Chipman, D. M. (1992) Degradation of Bacillus thuringiensis ssp. israelensis δ-endotoxin in vitro and in mosquito larval gut. Biocontrol Sci. Technol. (in press).Google Scholar
Pendleton, I. R. and Morrison, R. B. (1966) Separation of the spores andcrystals of Bacillus thuringiensis. Nature 212, 728729.CrossRefGoogle Scholar
Sommerville, H. J. (1978) Insect toxin in spores and protein crystal of Bacillus thuringiensis. Trends Biochem. Sci. 60, 181197.Google Scholar
Thomas, W. E. and Ellar, D. J. (1983) Bacillus thuringiensis var israelensis crystal δ-endotoxin effects on insect and mammalian cells in vitro and in vivo. J. Cell Sci. 60, 181197.CrossRefGoogle ScholarPubMed