Analysis Of Antifungal Active Ingredients From Streptomyces Js-1~T And Its Biological Activity Determination

In this thesis we mainly research the metabolite properties identification of the strains Streptomyces Js-1T,purification and structure identification of antifungal active substances from Streptomyces Js-1T metabolite and the antibacterial activity test of it.Properties of Streptomyces Js-1T metabolites show,Js-1T has wide antifungal spectrum,all selected pathogenic fungi strains has been effect inhibitory by fermentation broth of Js-1T.It manifestation the most obvious inhibitory effect to Mcogone perniciosa and it antifungal rate was 100%.Js-1 fermentation broth can valid suppress the strains Phomopsis longanae,Botrytis paeoniae and Hypomyces aurantius,ihe antifungal rate was above 60%to those strains.Js-1T fermentation broth antifungal rate range 30%to 50%for the strains Lasiodiplodia theobromae,Rhizopus,Trichodermaviride and Aspergillus niger,the antifungal rate range 20%to 25%for the strains Phyricularia grisea and Hypoxylon sp,the antifungal rate at around 10%for the strains Pestalotiopsis theae,Fusarium graminearum,Fusarium oxysporu f.sp and Colletotrichum frigidium.But the antifungal activities of the mycelia extracts were detected that antifungal rate under 40%whole,it below the Fermentation broth by inhibiting all selected pathogenic fungi strains and it show no inhibition in grows of Colletotrichum frigidium and Fusarium graminearum.The antibacterial experiments show that Js-1T fermentation broth has distinct inhibition to gram-positive and gram-negative bacteria,but no inhibition to Saccharomyces cerevisiae.The other metabolite properties identification indicate that the product after pretreatment fermented broth were very stability of pH,temperature,ultraviolet and storage time.Only n-butyl can be extracted the antifungal activities matter effectively of all test dissolvent,which show that the antifungal activities matter has hight polarity.However compever with the n-butyl the D101 macroporous adsorption resin can isolate active substances more effectively.The adsorption characteristics of antifungal active substances by D101 macroporous adsorption revealed:the best static saturated adsorption capacity of active substances for D101 macroporous adsorption is 25 g/200 mL,the time of static adsorption saturation is 3 h and the best solution of adsorbent for distilled water.The product from retreatment of fermentation broth has been Separated and purificatory by the Separation methods with D101 macroporous adsorption resin,silica-gel column chromatography,C18 Reverse phase silica gel column chromatograph and Sephadex LH-20 chromatographic column,after those methods we obtain a pure antifungal activity substance with 95.52%purity,it was named JS-B2.JS-B2 was analyzed by HPLC,ESI-MS and NMR,the results display that JS-B2 is a new structure compound,and it molecular weight is 167,molecular Formula is C9H13NO2,English name of structural formula is(2E,6Z)-2-aminocycloocta-2,6-dienecarboxylic acid.Under the action of JS-B2 the lethal concentration of 50%(Lc50)of Mcogone perniciosa is 1.77μg/mL and it lethal concentration of 90%(Lc90)is 2.88μg/mL,but the Lc50 of Agaricus bisporus is 3.26μg/mL and it Lc90 is 7.14μg/mL.The Lc50 and Lc90 of Hypomyces aurantius were 3.09 μg/mL and 5.26μg/mL,but the Lc50 and Lc90 of Hypsizygus marmoreus were 8.37 μg/mL and 122.62 μg/mL.Antifungal activity result show that compare with antifungal drug nystatin and miconazole JS-B2 has better inhibition pathogenic fungi grows.Respectively antifungal rate were 100%,100%,92.78%,80.56%,58.33%of Mcogone perniciosa,Hypomyces aurantius,Phomopsis longanae,Aspergillus niger and Lasiodiplodia theobromae.Therefore,it has great application value of the antifungal active substances JS-B2.