Separation And Purification Of Tautomycin And The Anti-fungal Activity Of The Compounds With Maleic Anhydride Structure

Tautomycin is an antibiotic produced by Streptomyces spiroverticillatus. It has a strong activity in inhibiting the growth of Sclerotinia sclerotiorum and Botrytis cinerea, as well as the growth of many kinds of fungi. Moreover, tautomycin is a kind of protein phosphatase inhibitors, displaying strong inhibition against both protein phosphatase-1 and protein phosphatase-2A, So tautomycin has great potential in practical use. In this paper, methods of HPLC detection of Tautomycin, separation and purification of tautomycin from fermentation broth, the bacteriostatic activity of tautomycin and related compounds containing the structure of maleic anhydride against fungi are studied.Firstly, a high performance liquid chromatography (HPLC) method for detection of tautomycin in broth was established. Pretreatment was essential to convert tautomycin into its corresponding disodium salt. The the optimized HPLC conditions was as follows: the mobile phase consisting of water-acetonitrile (60:40), wavelength of detection 254nm, flow rate of mobile phase 1.0 ml/min, the retention time: 2.5 min. The precision study showed that the method was efficient and precise enough to measure the concentration of tautomycin in samples.A separation and purification process of tautomycin from fermentation broth was also described. Tautomycin sample was acquired through several steps including filtration, immersion, ion exchange, extraction, and silica gel column chromatography. And the specific conditions of each step have been explored and compared. The total recovery is about 20%- 30%.The inhibition activities of tautomycin and other compounds containing the structure of maleic anhydride against Aspergillus niger and Penicillium sp. were determined by means of oxford cup. The result demonstrated that tautomycin had strong antimicrobial activity mainly because of its side-chain. The introduction of methyl group, chlorine and bromine atom to the side-chain of the maleic anhydride could its increase inhibition activity, while the introduction of phenyl group and fluorine atom showed negative effect on its antimicrobial activity.