三株海洋微生物活性次级代谢产物研究

发布时间：2018-05-14 02:08

  本文选题：海洋链霉菌 + 深海真菌　； 参考：《浙江大学》2017年博士论文

【摘要】：本文对从温州洞头近海沉积物样品和南海深海沉积物样品中分离得到的121株菌的发酵粗提物进行了细胞毒活性筛选,选择其中两株具有显著细胞毒活性的菌株Streptomyces sp. DT-A37和Penicili m sp. NH-SL 以及 1 株代谢产物类型丰富的菌株Alternariasp.NH-F6进行代谢产物系统分析,共分离鉴定了 46个化合物,其中9个为新化合物,并对所得化合物进行了细胞毒活性测试、BRD4蛋白抑制活性测试、抗菌活性测试等生物活性评价。综合运用常压硅胶柱色谱、反相高效制备液相色谱、凝胶柱色谱等方法对发酵提取物进行分离纯化,采用波谱技术鉴定化合物结构。运用OneStrain-Many Compounds (OSMAC)策略对链霉菌Streptomycessp. DT-A37的代谢产物进行挖掘,从中分离鉴定了 10个化合物,包括4个新化合物(1Z)-S,S'-dimethyldihydro holomycin (2), holomycin A (3), cis-2'- (4"-hydroxy-4"-methylpenty1) - cyclo-propylacetic acid (4), cis-2'- (4", 5"-dihydroxy-4"-methylhexyl) -cyclopropylaceti acid (5),以及6个已知化合物。其中,holomycin (1)具有显著的细胞毒活性和抗菌活性,其对H1975细胞的IC50值为1μM,对大肠杆菌和金黄色葡萄球菌的MIC值均为64 μM。从交链孢霉菌Alternariasp. NH-F6的固体发酵培养提取物中共分离鉴定了26 个化合物,包括 5 个新化合物 8β-chloro-3, 6aa, 7β, 9β,10-pentahydroxy-6, 6a,6b, 7, 8, 9-hexahydro-4 (5H) -one (11), 3, 7β, 8α, 9β, 10-pentahydroxy-7, 8-dihydroperylen-4 (9H) -one (12), 2- (N-vinylacetamide) -3-ene-4-hydroxymethy1-y-butyrolactone (13),alternarienonic acid B (15), chrysogeside F (17),以及 21个已知化合物。其中,化合物11和12具有显著的BRD4蛋白抑制活性,IC50值分别为11.8 μM和7.1 μM。化合物12具有明显的抗菌活性,其对大肠杆菌和金黄色葡萄球菌的MIC值分别为16 μM和32 μM。从青霉菌Penicilliumsp. NH-SL的液体发酵培养提取物中共分离鉴定了 10个化合物,其中具有异戊烯基取代的吲哚生物碱类化合物verrucofortine (38)表现出显著的细胞毒活性,其在测试浓度为0.7 μM时对Hepa lclc7细胞的抑制率为85.8 %。采用多级质谱法对该化合物进行裂解规律的研究,并根据其裂解规律,从该菌株的总提取物中检测到其它三个异戊烯基吲哚生物碱类似物47～49,为异戊烯基吲哚生物碱类化合物的快速发现提供了参考依据。
[Abstract]:In this paper, the fermentation crude extracts of 121 strains isolated from Dongtou offshore sediment samples and deep-sea sediment samples from the South China Sea were screened for cytotoxicity, and two of them, Streptomyces spp., were selected. DT-A37 and Penicili m sp. NH-SL and Alternariasp.NH-F6, a strain with abundant metabolites, were analyzed systematically. A total of 46 compounds were isolated and identified, 9 of which were new compounds. The cytotoxic activity of the compounds was evaluated by BRD4 protein inhibition test and antimicrobial activity test. The fermentation extract was separated and purified by atmospheric pressure silica gel column chromatography, reversed phase high performance liquid chromatography and gel column chromatography. The structure of the compound was identified by spectroscopic technique. Streptomyces spp. Ten compounds were isolated and identified from the metabolites of DT-A37, including four new compounds, namely, four new compounds, one new compound, one, two, four, five, four, four Its IC50 value to H1975 cells was 1 渭 M, and the MIC value to Escherichia coli and Staphylococcus aureus was 64 渭 M. Alternaria sp. from Alternaria alternata. NH-F6鐨勫浐浣撳彂閰靛煿鍏绘彁鍙栫墿涓�鍏卞垎绂婚壌瀹氫海�

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