Separation, Purification, And Identification Of Antibiotic Product Of Streptomyces Lydicus A02

Antigonstic strain A02 is the streptomyces lydicus which is isolated by our laboratory and ithave a high antimicrobial activity. This paper has a systemic analyzed separate extraction andpurification method, physical-chemical character, the structure and the toxicity of antibotic ofStreptomyces lydicus A02.The result as follows:1 Analyzed the physical-chemical character of fermentation broth of antigonistic strain A02.Fermentation broth A02 strain displays a higher stability to heat and acid. It could keep a goodantifungal activity under the conditions of temperature below 70℃,pH5～8; The storage stabilityof A02 fermentation broth is good under 4℃, but under the hot conditions activeness loses was big;The A02 fermentation broth is stable under the daylight lamp and the natural light, but it issensitive to the ultraviolet ray; By the polarity test, we knowed that the active component of A02fermentation broth is neutral and the multi-alkenes antibiotic.2 Studied the extraction and purification method of active component of A02 antibioticproduct, and use of the method we obtained the pure of A02 antibiotic product with high-purity.The methods of Separation and purification is as follows:â‘ pretreatrnent of fermentation broth;â‘¡Using macroporous absorption resin: fermentation broth (1:1,v/v) to, first, using the doublevolume non-ion water to elution, second, using the double volume 30% methanol-water(3:7,v/v) toelution, at last, using the double volume ethanol-water(7:3.v/v) to elution, collecting activecomponent, and then concentrating it.â‘¢Using theconcentrated solution to full with Silica gelcolumn, and selecting the solvent system of ethanol-ammonia-water(8:1:1,v/v) to elution,collecting active component, and then concentrating it.â‘£Using the concentrated solution to fullwith HPLC, with the mobile phase of methanol-water(7:3,v/v), collecting active component, andthen concentrating it.⑤The concentrated solution puts in 45℃constant temperature to dry oven,and finally we obtained the cream color solid powder, and the purity is 99.845%。3 Determined the A02 antibiotic product pure structure, and clear knowed about the puremolecular formula and the structural forrnula.The use of the ultra-violet spectrograph to determine structure of A02 antibiotic product, theresult showed that A02 pure is four alkenes agricultural antibiotic; By analysis of infrared spectrum result indicated, A02 antibiotic product pure includes expansion and contraction vibration of-OH,-NH₂,-CH₃,-CH₂,-C=O,-C=C-and so on; The use of mass spectrum to determine the A02 puremolecular weight is 665, the molecular formula is C₃₃H₄₇NO₁₃; Seeing from the 13C spectrum, A02antibiotic product pure molecule includes the carboxyl group carbon atom, not the saturated fourcarbon atoms, on the hexose five carbon atoms, the methyl carbon atom, with amino connectedcarbon atom; Seeing from the 1H spectrum, A02 antibiotic product pure molecule includes protonhydrogens which is on the not the saturated four carbon atoms, and in carbon connection oxygenproton hydrogen, with hydroxyl connection proton hydrogen. Through above spectral analysis,finally promotes A02 antibiotic product chemistry structural。4 Studied the A02 antibiotic product pure solubility and the functional group response.A02 antibiotic product pure may dissolve in the methyl alcohol, DMSO, acetic acid,Dissolves slightly in the trimethylene glycol, the normal butyl alcohol。Extremely micro dissolutionin ethanol; Difficult to dissolve in the water。The A02 antibiotic product pure functional groupresponded show that A02 antibiotic product pure includes sugar glucoside key, amino acid uncleamino and aldehyde group and does not contain the return to original state sugar and the phenolicgroup。Responded with the potassium permanganate and the strong sulfuric acid that A02 antibioticproduct pure has he the similar structure with the thin large ring lactone class antibiotic.5 Bioassay of toxicity of A02 antibiotic product, and clear knowed about its EC₅₀ and EC₉₅The bioassayed result indicated that the A02 product has good inhibitory effect to the Botrytiscinerea, Its EC₅₀ and EC₉₅ value respectively is 2.9373 mg/L and 53.0919 mg/L, and control effectobviously goodness than the comparison the chemical fungicide Lide(EC₅₀ and EC₉₅valuerespectively is 7.6245 mg/L and 277.6080mg/L).