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Combining spectroscopy method has been used to kill fungal drug screening and molecular analysis of the interaction, but Since fungi CYP51 (lanosterol 14α-demethylase) is a membrane-bound proteins, it is difficult purified to obtain a sufficient amount and activity of the target enzyme Therefore, usually in studies using purified soluble prokaryotic CYP51 protein, this approach will inevitably cause errors. Establish a fast and accurate method for the screening of the target enzyme CYP51 fungicides, has important significance to the mass screening of a new synthetic drug. Use this method to analyze the binding capacity of the fungicides and the target enzyme CYP51 screening potential application value of new fungicides and to optimize the design and synthesis of highly efficient, low toxicity and high selectivity DMIs fungicide compounds provide an important basis for . Rice blast fungus to rice blast fungus microsomes and heterologous expression of CYP51, analysis of tebuconazole allyl compounds, tebuconazole, triazole alcohol triadimefon on behalf of the the DMIs class to kill the fungal agent CYP51 binding capacity, Analysis of the impact of the target activity, the purity of the target enzyme, and the concentration of the target enzyme combination spectrum analysis and comparison with the results of the biological tests for its reliability. The results show that the activity of the target enzyme, no the other P450 interference and a suitable concentration of the target enzyme is a necessary condition to obtain accurate binding spectrum. Diniconazole, tebuconazole, triadimenol, triadimefon target enzyme binding constant (Kd) for 0.143μM 0.24μM 0.257μM, 0.307μM, and the results of its rice blast fungus growth inhibition (120h- EC 50 ) was significantly related to to prove spectrometry can be used as a simple and reliable DMIs class fungicide screening methods. In E. coli expression CYP51 as material, the different structures of the drug with the binding ability of the CYP51 results show that: the the ring drugs EVE-2 containing indole-3 EVE and LHEXP-10, LHEXP-3s are combined and CYP51 combined constants were 73.039μM, 18.634μM, 2.18μM, 66.998μM; triazole imidazoline TZDs FBQbp-25 combined with the CYP51, Kd value 1.607μM of, FBQbp-57 and CYP51; imidazolinone drugs The SYL-25 SYL-35, SYL-36, DM-2, SC-11 with CYP51 closely integrated, Kd values ??for 5.147μM 8.653μM 10.199μM, 3.603μM and 3.795μM, while SYL-27 DM-5, SC-17, SYC-4 and CYP51 completely combined, SYL-26, with Syl-33, SYC-11 and ZD-6 and CYP51 part of the combination, but the combination of the lower specificity. The study also found part of the drug and the binding capacity of the CYP51 growth inhibition test results do not conform with rice blast fungus. The same types of drugs, the number of the substituents in the case of the same basic structure, location, and the different types, so that they are different and the binding ability of CYP51. 19 kinds of drug screening results, wherein EVE-3, LHEXP-10, FBQbo-25, SYL-25, SYL-35, SYL-36, DM-2, SC-11 having a bactericidal potential.
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