Studies On The Metabolites Of Amycolatopsis Orientalis HCCB10007

Vancomycin, a glycopeptide antibiotic, has been used as the first-line drug for treating serious MRSA infections, and its reseach is a hot topic in the world. With the application and thorough study of vancomycin, the analysis of effective components and related compounds are increasing more and more attention. Domestic vancomycin product has been commecialized already. However its related compounds have not been studied yet. With vancomycin-resistant bacteria has been increasing, people are afraid that vancomycin will not effective persistently. Under this circumstance, it is necessary to carry research and development on novel glycopeptide antibiotics for treating serious infections.This paper is attempt to establish purification, analysis and identification methods of domestic vancomycin related compounds. Also studied the influences of different fermentation conditions to these components in the metabolites of strain Amycolatopsis orientalis HCCB10007. To gain fermentation media which to product different production, also to optimize and utilize the fermentation media, to lay the groundwork for further studies of glycopeptide antibiotics.The main achievements:(1) Related compounds were deeply studied, a specific macroporous adsorptive resin was screened for isolation process. Combined with other methods like medium-pressure LC method and HPLC preparation method, seven compounds were obtained from the fermentation broth and crude product. Five of them were identified, through MS and chemical analysis, as: Demethylvancomycin B,Desamidovancomycin B₁ and B₂, Desvancosaminylvancomycin B, Aglucovancomycin B. All of them are analogues of vancomycin. Among them, desamidovancomycin B₁ and B₂ are a pair of isomers, which remains a certain ratio in solvent, The other two compounds had molecular wieght at 1318D and 1459D. Studies the physical and chemical properties of these compounds, the activity of Aglucovancomycin B in vitro is higher than Vancomycin B.(2) The changes of metabolites in different fermentation conditions, especially the nitrogen sources, of Amycolatopsis orientalis HCCB10007 were examed. Three fermentation media were found with significant changes on yield and rate of vancomycin and other components: a synthetic media with vancomycin percentageabove 70%, while other components greatly decreased; a vancomycin high producing yeast media; and a media that could produce didechlorovancomycin. The first two media were optimized for future potential application. Didechlorovancomycin was purified and identified, and its production conditions was studied. This compound may be used as new identity for further synthetic use.(3) Synthetic media were used to the fermentation test of genetic engineering bacteria HCCB10072 and HCCB10082, fermentation results is better than the control medium. To obtained two aglycones of vancomycin from fermentation method. To enlarge the experiments of yeast media in 5L fermentor, the yield of vancomycin is higher than control. And also done the cyclical alternative experiment, initially study the feasibility for re-use of vancomycin waste residue.