title: Endogenous protease-activated 66-kDa toxin from Bacillus thuringiensis subsp. kurstaki active against Spodoptera littoralis.
abstract: The anti-lepidopteran toxin from sporulated Bacillus thuringiensis subsp. kurstaki cells, generated by the proteolytic action of endogenous protease(s) on the protoxin, was purified and studied to identify the effect of such proteolysis on the biochemical nature of the toxin. The active toxin was purified employing anion-exchange chromatography to absolute homogeneity, as indicated by SDS-PAGE and Western blotting. Antisera to the purified toxin (66 kDa) crossreacted with the protoxin (132 kDa) confirming its origin from protoxin. The purified toxin with a pI of 7.95 was derived from the N-terminal region of the protoxin (pI 7.6). Circular dichroism data revealed that the toxin has significant secondary structure and it undergoes pH dependent conformational change. Unlike the toxin generated by exogenous proteases such as trypsin, etc., the endogenous protease(s) activated toxin is highly lethal to a tolerant insect variety of the lepidopteran order, Spodoptera littoralis.
authors.affiliation: Department of Biochemistry, Osmania University, Hyderabad, India.
authors.forename: N S
authors.initials: NS
authors.lastname: Kumar
authors.forename: G
authors.initials: G
authors.lastname: Venkateswerlu
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001413:Bacillus thuringiensis / Q000737:chemistry* / Q000201:enzymology*; D001427:Bacterial Toxins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D001681:Biological Assay; D015153:Blotting, Western; D002942:Circular Dichroism; D010450:Endopeptidases / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D011498:Protein Precursors / Q000302:isolation & purification; D017433:Protein Structure, Secondary; D013045:Species Specificity; D018411:Spodoptera / Q000382:microbiology*
chemical_list: D000596:Amino Acids; D001427:Bacterial Toxins; D011498:Protein Precursors; D010450:Endopeptidases