[Purification and partial characterization of antagonistic proteins from Bacillus subtilis B034]

Bacillus subtilis B034, an antagonistic bacteria isolated from the phyllosphere of rice, strongly inhibits the growth of Xanthomonas oryzae pv. oryzae which causes the rice bacterial blight. Crude extract was obtained by precipitation of the cell-free culture of B034 with 70% saturated (NH4)2SO4. The suspension of the precipitate strongly inhibits Xanthomonas oryzae pv. o-ryzae, and it is thermostable, resistant to trypsin, partial sensitive to proteinase K, pronase E and chloroform. Its active pH range is wide, from 4.0 to more than 12.0, relative more stable in high pH. Two antagonistic peaks were obtained from the crude extract of B034 after Phenyl-Sepharose CL-4B, DEAE-Sephacel and FPLC Superdex 75 HR 10/30 column chromatography. One of these two peaks, named P2, was showed only single band with 50.3 kD and pI6.25 in SDS-PAGE and PAGE-IEF, respectively. The N-terminal partial sequence of protein P2 was analyzed.