Studies On Purification And Properties Of Antagonistic Protein From Endophytic Bacteria SS02 Of Paris Polyphylla Var.Chinensis

In this thesis, a broad-spectrum antibiotic Endophytic strain was screened outby antifungal exogenous experiments, basing on seperating Endophytic BacteriaSS02 of Paris Polyphylla Var. Chinensis comprehensively, then it was named SS02after molecular identifying.SS02 was incubated through different temperature, time and PH, and theantibacterial competence was checked, then an conclusion was come to: the bestcondition of fermenting the protein is incubating it in PB at 30℃, 7.0PH, shaking it72 hours in bassinet and purifying it throughing on 20%-50%(NH₄)₂SO₄.The zymolytic liquor was putted in centrifuge, 10 min lasted and12000rpm, and the residua and thalli was cleared. After purified throughing on(NH₄)₂SO₄, the antibacterial competence of zymolytic liquor was gone, andsynchronously, the obvious antibacterial competence was detected indeposit. When the deposit was gathered after centrifuge and it was dissolved inphosphate buffer, the original protein was gained. Then a antibacterial protein wasseperated when different protein was gathered through purifying original protein ingel pole and fibre pole respectively and contrasted against plant pathogen by trackexperiment. The SDS-PAGE shown that the protein is pure. Its molwt wasmensurated—56000 and a single stripe was shown in electrophoresis. Its PI valueis 6.4 in PAGE-IEF, it is thermostabale, stable in ultraviolet and partially sensitiveto enzyme. Purified SD22 can strongly restrain strains such as Rhizoctonia cerealis,Sclerotinia sclerotiorum Physalospora piricala, Trichodema viride, Gliocladiumviride, Curvularia leaf spot, Fusarium sp, Fusarium head blight, BeauveriaBassiana, Escherichia coli, Staphylo-coccus aureus, Bacillus subtilis, Candidalvaginitis, Fusarium oxysporum Schl.emend.Snyder&Hansem.