Study On Hydroxylation Epoxyprogesterone Marine Fungus Strain Screening And Metabolites

Adrenal cortical hormone drugs as an important part of the steroid hormone drug, is widely used in clinical.The introduction of the C11-hydroxyl group is a important conversion reaction of steroidal.The method of introducing the main C11-hydroxyl group including chemical and biological transformation methods. Biotransformation method has the advantage of shortcutthe and environment friendly,so this method is widely used.In this thesis, we use epoxyprogesterone as the substrate,screening the active strains among eigh marine fungus. We expect we can acquire a new biotransformation technology of steroid hormone drug.Considering the unique growth environment of marine fungi, we use marine fungi MNP07010101as the research object. By measuring the activity of its mycelia and fermentation broth extract,we determine the polarity of the active part of the region.Thus effectively guiding the separation of marine fungal metabolites, and finally we get the marine fungus metabolite having biological activity(1) By thin layer chromatography, high performance liquid chromatography and liquid chromatography mass spectrometry to study the situation of epoxyprogesterone conversion by the eight marine fungus.Finally, we filter out the active strain that the marine fungi MNP07010101has the11-hydroxylation capacity.We determine the marine fungi MNP07010101as the ocean chrysogenum(Penicillium chrysogenum) and the strain has theC11-hydroxylation capacity for the first time reported.(2) The marine fungi MNP07010101fermentation broth followed by ethyl acetate extraction, high performance liquid chromatography analysis,semi-preparative liquid chromatography,six transformation products were found.Through structural characterization, two of them were identified as11α-hydroxy epoxyprogesterone and11β-hydroxy epoxyprogesterone.(3) We optimize the conversion process of11α-hydroxy-epoxy progesterone.We establish the best medium components as follows: glucose was3%, corn starch was2.5%, ammonium sulphate was0.16%the leaching yeast powder0.08%; fermented liquid initial optimal pH value was5, the optimal feeding time is36h, optimum inoculation quantity was6%, the optimum table speed of180·min-1, the best time for36h and established the3%ethanol suspension feeding is the most appropriate way of feeding.(4) Culture conditions of marine fungi MNP07010101were optimized to establish the malt the medium (sea water:water=2:8) and cultured for21days was the suitable culture conditions, and then mass culture,we preparate of the mycelium and fermentation broth extract.(5) Through the DPPH radical scavenging ability test, the trypsin inhibitory experiment and anti-tumor activity experimental, we found that the marine fungus MNP07010101has good tumor suppressor activity, and the preferably tumor suppressor activity of this strain was reported for the first time.Activity tracking method is then used to determine the anti-tumor activity site.(6) By column chromatographic techniques for the separation of active secondary metabolites of the anti-tumor activity site. The two compounds were found. One of them was identified as the peroxygen ergosterol, i.e.5a,8a-peroxy ergost-6,22E-diene-3β-ol, and that the compound obtained from the strains was isolated for the first time.