海洋长孢葡萄穗霉菌生物合成纤溶活性化合物的研究

Research of biosynthesis of fibrinolytic compounds from marine Stachybotrys longispora

目的基于FG216(Fungus stachytotrys longispora)产生纤溶化合物FGFC1(Fungi fibrinolitic compound 1)的可能生源途径,调控其次生代谢过程,获得新型纤溶活性或者其他活性化合物。方法 FG216经过种子培养和发酵培养,在发酵培养结束前24h,添加ADP、色氨酸、柠檬酸三钠、赖氨酸、苯丙氨酸、组氨酸、鸟氨酸、天冬酰胺等8种代谢产物调节剂(metabolite regulator,MTRL),添加量为发酵液的1%,用HPLC(High-pressure liquid chromatogram)检测FG216代谢产物产出情况。结果添加鸟氨酸为MTRL的FGFC1的产量增加至950mg/L,苯丙氨酸为MTRL的FG216代谢产物的HPLC图谱出现2个新的化合物峰。色氨酸为MTRL的FG216代谢产物的HPLC图谱出现3个新的化合物峰。ADP、柠檬酸三钠、赖氨酸、组氨酸、天冬酰胺为MTRL的FG216代谢产物的HPLC图谱未出现新的化合物峰。分离纯化5种目标化合物CA、CB、CC、CD、CE在纤溶活性评价反应体系(10μg/mL),其纤溶活性分别是FGFC1的30%、90%、50%、110%、17%。结论采用恰当的FG216发酵MTRL,能产生新型的纤溶活性化合物,推测通过调控代谢生源途径,能从FG216挖掘出活性更强或结构新颖的纤溶化合物。

Objective To regulate secondary metabolic processes of the strain FG216 and obtain novel compounds with fibrinolytic activity or other activities based on the possible biosynthetic pathway of fibrinolytic compound FGFC1 produced by FG216.Methods FG216 was seeded and fermented,then ADP,tryptophan,trisodium citrate,lysine,phenylalanine,histidine,ornithine,asparagine were added 24 hours before the end of fermentation.A metabolite regulator(MTRL)was added in an amount of1% of the fermentation broth,and the production of metabolites of FG216 was detected by HPLC.Results The production of FGFC1 increased to 950 mg/L when MTRL was ornithine.HPLC chromatograms of the metabolites of FG216 with phenylalanine as MTRL showed peaks of two new compounds.HPLC chromatograms of the metabolites of FG216 with tryptophan as MTRL showed peaks of three new compounds.HPLC chromatograms of the metabolites of FG216 with ADP,trisodium citrate,lysine,histidine,and asparagine as MTRL showed no peaks of new compounds.The target compounds CA,CB,CC,CD,CE in the fibrinolytic activity evaluation system(10μg/mL)were isolated and purified,and their fibrinolytic activities were 30%,90%,50%,110%,and 17% of fibrinolytic activity of FGFC1,respectively.Conclusion The use of the appropriate MTRL in fermentation of FG216 can produce new fibrinolytic compounds.It’s presumed that fibrinolytic compounds with stronger activity or novel structure can be excavated from FG216 by regulating the metabolic pathway.