Purification of the insecticidal toxin in crystals of Bacillus thuringiensis.

Crystals were purified from four serotypes of the insect pathogen Bacillus thuringiensis. Crystals from these serotypes were similar in amino acid and N-terminal analyses, but differed in their toxicity to two species of Lepidoptera and in their immunological properties. Toxic polypeptides were obtained following trypsin digestion of solutions of the crystals. In two strains (serotypes 3 and 9) this fraction contained only one polypeptide. Similar results were obtained when dissolved crystals were digested with other proteolytic enzymes or with gut contents from Pieris brassicae. The trypsin-resistant polypeptide was further purified by gel and ion-exchange chromatograhy and had a molecular weight of about 70,000, estimated by gel chromatogrpahy and gel electrophoresis. No evidence was obtained for a toxin of lower molecular weight. This purified toxin accounted for most, if not all, of the toxic activity originally present in the crystal solution and was active by injection and ingestion. The purified toxic fraction from serotype 1 appeared to contain two polypeptides, one of which corresponded to that found with serotypes 3 and 9. There were no major differences in the composition of crystals from different serotypes of B. thuringiensis and it is concluded that the trypsin-resistant polypeptide represents the active insecticidal toxin of the crystal.