Characterization of larvicidal toxin protein from Bacillus thuringiensis serovar japonensis strain Buibui specific for scarabaeid beetles

The delta-endo toxin proteins from Bacillus thuringiensis which kill the larvae of various scarabaeid beetles such as Anomala cuprea, A. rufocuprea and Popillia japonica were purified by DEAE ion exchange chromatography. A protein with a molecular size of 130 kDa was purified. During the purification a minor peak was also detected which was estimated to be 67 kDa by SDS-PAGE. Both 130 and 67 kDa proteins showed larvicidal activity against A. cuprea. The lethal concentration of the 130 kDa protein which killed 50% of the larvae tested (LC50) against A. cuprea was 2 micrograms g-1 compost. A comparison by SDS-PAGE of the V8 protease digestion pattern of the 130 and 67 kDa larvicidal proteins showed that proteolytic resistant core peptides of approximately 60 kDa molecular size were resulted. The N-terminus amino acid sequence of the 130 and 67 kDa proteins was determined to be NH2-XXPNNQNEYEIIDAL and NH2-XSRNPGTFI, respectively, which is not identical to the sequence of CryIA, CryIB, CryIC and CryIII proteins.