两株抗肿瘤土壤放线菌代谢产物的分离鉴定
发布时间：2019-05-20 14:16
【摘要】：抗肿瘤抗生素在临床中广泛使用,但存在药物毒副作用及肿瘤细胞多耐药性等问题,为了解决这一难题,寻找结构新颖的抗肿瘤化合物,本研究采用活性追踪的方法开展了抗肿瘤土壤放线菌筛选及对两株活性放线菌次级代谢产物的研究工作。以人肺癌细胞A549和人结肠癌细胞HCT-116为筛选模型,采用CCK-8方法从土壤放线菌中筛选出两株活性菌,经16S r RNA初步鉴定均为链霉菌,分别命名为Streptomyces sp.HS-NF-1006和Streptomyces sp.HS-NF-853。结合抗肿瘤模型筛选结果和菌株在不同培养基中的生长状况,最终选定葡萄糖1%,可溶性淀粉4%,酵母抽提物0.4%,麦芽提取物0.1%,CaCO_3 0.2%,FeSO_4·7H_2O 0.1%,MnCl_2·4H_2O 0.1%为Streptomyces sp.HS-NF-1006的最适发酵培养基;选定葡萄糖1%,可溶性淀粉4%,酵母抽提物0.4%,麦芽糊精1%,0.1 ml/L(Fe SO_4·7H2O 0.1%,MnCl_2·4H_2O 0.1%,ZnSO_4·7H_2O 0.1%)为Streptomyces sp.HS-NF-853的最适发酵培养基。利用50 L发酵罐分别对两株活性菌进行放大发酵。所得发酵液经前处理后,利用硅胶、凝胶、HPLC等分离方法对两株活性菌的发酵产物进行分离,最终得到9个化合物,利用核磁共振和质谱等分析方法进行结构鉴定,从菌株HS-NF-1006提取到3个化合物1-3,均为蒽环类化合物,其中化合物1为新结构化合物,命名为13-oxy-13-decarbony-6′′-cyano-6′′-deoxy-TAN-1120;从菌株HS-NF-853提取到6个化合物4-9,其中化合物4是一个结构新颖度较高的四氢苯并呋喃酮化合物,命名为Strefuranone A。对所得到化合物进行抗肿瘤活性评价,化合物1对结肠癌细胞HCT-116的IC50为0.0978μM,对人肺癌细胞A549的IC_50为0.08μM,化合物2-9未表现抗肿瘤活性。
[Abstract]:Antineoplastic antibiotics are widely used in clinic, but there are some problems, such as side effects of drugs and multi-drug resistance of tumor cells. In order to solve this problem, find novel antitumor compounds. In this study, the screening of actinomycetes in antitumor soil and the secondary metabolites of two strains of actinomycetes were studied by activity tracking method. Using human lung cancer cell line A549 and human colon cancer cell line HCT-116 as screening models, two strains of active bacteria were screened from soil actinomycetes by CCK-8. They were identified as Streptomyces by 16s r RNA. Named Streptomyces sp.HS-NF-1006 and Streptomyces sp.HS-NF-853., respectively. Combined with the screening results of antitumor model and the growth of the strain in different media, glucose 1%, soluble starch 4%, yeast extract 0.4%, malt extract 0.1%, CaCO3 0.2% were selected. FeSO_ 4.7H _ 2O 0.1%, MnCl2 路4H _ 2O 0.1% is the optimum fermentation medium for Streptomyces sp.HS-NF-1006. Glucose 1%, soluble starch 4%, yeast extract 0.4%, maltodextrin 1%, 0.1 ml/L (Fe SO_ 4 路7H2O 0.1%, MnCl2 路4H2O 0.1%), ZnSO_ 4.7H _ 2O 0.1%) was the most suitable fermentation medium for Streptomyces sp.HS-NF-853. Two strains of active bacteria were amplified and fermented in 50 L fermenter. After pretreatment, the fermentation products of the two active bacteria were separated by silica gel, gel and HPLC, and nine compounds were obtained. The structure of the fermentation broth was identified by nuclear magnetic resonance (NMR) and mass spectrometry (MS). Three compounds 1 / 3 were extracted from strain HS-NF-1006, all of which were anthracycline compounds, in which compound 1 was a new structural compound named as 13 脳 13 / decarbony / 6 / cyano / 6 / deoxyto1120; Six compounds 4 鈮，
本文编号：2481703
[Abstract]:Antineoplastic antibiotics are widely used in clinic, but there are some problems, such as side effects of drugs and multi-drug resistance of tumor cells. In order to solve this problem, find novel antitumor compounds. In this study, the screening of actinomycetes in antitumor soil and the secondary metabolites of two strains of actinomycetes were studied by activity tracking method. Using human lung cancer cell line A549 and human colon cancer cell line HCT-116 as screening models, two strains of active bacteria were screened from soil actinomycetes by CCK-8. They were identified as Streptomyces by 16s r RNA. Named Streptomyces sp.HS-NF-1006 and Streptomyces sp.HS-NF-853., respectively. Combined with the screening results of antitumor model and the growth of the strain in different media, glucose 1%, soluble starch 4%, yeast extract 0.4%, malt extract 0.1%, CaCO3 0.2% were selected. FeSO_ 4.7H _ 2O 0.1%, MnCl2 路4H _ 2O 0.1% is the optimum fermentation medium for Streptomyces sp.HS-NF-1006. Glucose 1%, soluble starch 4%, yeast extract 0.4%, maltodextrin 1%, 0.1 ml/L (Fe SO_ 4 路7H2O 0.1%, MnCl2 路4H2O 0.1%), ZnSO_ 4.7H _ 2O 0.1%) was the most suitable fermentation medium for Streptomyces sp.HS-NF-853. Two strains of active bacteria were amplified and fermented in 50 L fermenter. After pretreatment, the fermentation products of the two active bacteria were separated by silica gel, gel and HPLC, and nine compounds were obtained. The structure of the fermentation broth was identified by nuclear magnetic resonance (NMR) and mass spectrometry (MS). Three compounds 1 / 3 were extracted from strain HS-NF-1006, all of which were anthracycline compounds, in which compound 1 was a new structural compound named as 13 脳 13 / decarbony / 6 / cyano / 6 / deoxyto1120; Six compounds 4 鈮，
本文编号：2481703
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