Abstract
Cell-free wall, membrane, and cytoplasmic fractions were prepared from Bacillus sphaericus 1593, which exhibited toxic activity against larvae of the mosquito Culex pipiens var. quinquefasciatus. Breakage of 12- to 14-h cells by sonication or French pressure cell yielded toxic material which could be assayed in a standard mosquito larva bioassay. When sporulating cells of strain 1593 were fractionated, the majority of the toxic activity was localized in the cell wall rather than in the plasma membrane or cytoplasm. The toxin located in the bacterial cell wall was relatively stable, in that activity was unaffected by treatment with trypsin, pronase, CHCl3-CH3OH-water, Triton X-100, 8 M urea (30 min), heat (80 degrees C, 12 min), sonication, refrigeration, lyophilization, or freezing. Activity was destroyed by boiling for 10 min or by 0.01 N NaOH. Only about 1.0% of the activity present in purified cell walls could be recovered by a 2-h extraction with 8 M urea or 3 M guanidine hydrochloride. A comparison of the toxicity of a cell-free cell wall fraction with that of a sample consisting entirely of heat-stable spores indicated that the spore preparation was about 10 times more active.
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- Davidson E. W., Singer S., Briggs J. D. Pathogenesis of Bacillus sphaericus strain SSII-1 infections in Culex pipiens quinquefasciatus (equal to C. pipiens fatigans) larvae. J Invertebr Pathol. 1975 Mar;25(2):179–184. doi: 10.1016/0022-2011(75)90066-x. [DOI] [PubMed] [Google Scholar]
- Frieben W. R., Duncan C. L. Homology between enterotoxin protein and spore structural protein in Clostridium perfringens type A. Eur J Biochem. 1973 Nov 15;39(2):393–401. doi: 10.1111/j.1432-1033.1973.tb03137.x. [DOI] [PubMed] [Google Scholar]
- Hungerer K. D., Tipper D. J. Cell wall polymers of Bacillus sphaericus 9602. I. Structure of the vegetative cell wall peptidoglycan. Biochemistry. 1969 Sep;8(9):3577–3587. doi: 10.1021/bi00837a013. [DOI] [PubMed] [Google Scholar]
- KELLENBERGER E., RYTER A., SECHAUD J. Electron microscope study of DNA-containing plasms. II. Vegetative and mature phage DNA as compared with normal bacterial nucleoids in different physiological states. J Biophys Biochem Cytol. 1958 Nov 25;4(6):671–678. doi: 10.1083/jcb.4.6.671. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kalb V. F., Jr, Bernlohr R. W. A new spectrophotometric assay for protein in cell extracts. Anal Biochem. 1977 Oct;82(2):362–371. doi: 10.1016/0003-2697(77)90173-7. [DOI] [PubMed] [Google Scholar]
- Kellen W. R., Clark T. B., Lindegren J. E., Ho B. C., Rogoff M. H., Singer S. Bacillus sphaericus Neide as a pathogen of mosquitoes. J Invertebr Pathol. 1965 Dec;7(4):442–448. doi: 10.1016/0022-2011(65)90120-5. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lecadet M. M., Chevrier G., Dedonder R. Analysis of a protein fraction in the spore coats of Bacillus thuringiensis. Comparison with crystal protein. Eur J Biochem. 1972 Feb 15;25(2):349–358. doi: 10.1111/j.1432-1033.1972.tb01703.x. [DOI] [PubMed] [Google Scholar]
- Myers P., Yousten A. A., Davidson E. W. Comparative studies of the mosquito-larval toxin of Bacillus sphaericus SSII-1 and 1593. Can J Microbiol. 1979 Nov;25(11):1227–1231. doi: 10.1139/m79-193. [DOI] [PubMed] [Google Scholar]
- Myers P., Yousten A. A. Toxic activity of Bacillus sphaericus SSII-1 for mosquito larvae. Infect Immun. 1978 Mar;19(3):1047–1053. doi: 10.1128/iai.19.3.1047-1053.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ramoska W. A., Singer S., Levy R. Bioassay of three strains of Bacillus sphaericus on field-collected mosquito larvae. J Invertebr Pathol. 1977 Sep;30(2):151–154. doi: 10.1016/0022-2011(77)90212-9. [DOI] [PubMed] [Google Scholar]
- Singer S. Insecticidal activity of recent bacterial isolates and their toxins against mosquito larvae. Nature. 1973 Jul 13;244(5411):110–111. doi: 10.1038/244110a0. [DOI] [PubMed] [Google Scholar]
- Somerville H. J., Delafield F. P., Rittenberg S. C. Urea-mercaptoethanol-soluble protein from spores of Bacillus thuringiensis and other species. J Bacteriol. 1970 Feb;101(2):551–560. doi: 10.1128/jb.101.2.551-560.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- TREVELYAN W. E., PROCTER D. P., HARRISON J. S. Detection of sugars on paper chromatograms. Nature. 1950 Sep 9;166(4219):444–445. doi: 10.1038/166444b0. [DOI] [PubMed] [Google Scholar]


