叶柄粘球菌STXZ77的分离鉴定及抗肿瘤活性

发布时间：2018-01-13 03:00

本文关键词：叶柄粘球菌STXZ77的分离鉴定及抗肿瘤活性　出处：《中国生物工程杂志》2016年11期 　论文类型：期刊论文

【摘要】：目的:从土样中分离纯化粘细菌,对其进行鉴定与归类,以丰富粘细菌菌种资源,并对其进行抗肿瘤活性初步研究,为抗肿瘤药物开发奠定基础。方法:采用灭活大肠杆菌诱导法,从土样中分离纯化粘细菌,结合形态观察、生理生化特征及16S rRNA基因序列同源性分析进行菌株鉴定;向发酵液上清中加入XAD-16大孔吸附树脂提取发酵产物粗提物,CCK-8法进行体外抗肿瘤活性研究;RP-HPLC分离抗肿瘤活性组分,LC-MS/MS分析其分子质量。结果:分离并鉴定了STXZ77菌株,命名为Myxococcus stipitatus STXZ77。该菌株的发酵产物XAD-16树脂粗提物对小鼠黑色素瘤细胞B16、小鼠乳腺癌细胞4T1、人肝癌细胞SMMC-7721、人宫颈癌细胞He La、人结肠癌细胞SW480等多种肿瘤细胞具有较好的细胞毒性,作用24h的IC_(50)值分别为5.34μg/ml、13.50μg/ml、11.93μg/ml、28.70μg/ml、48.09μg/ml,而对正常细胞人脐静脉血管内皮细胞HUVEC的毒性较小,IC_(50)值为17.09μg/ml,小于B16、4T1及SMMC-7721的细胞毒性。RP-HPLC分离得到抗肿瘤活性组分AP-C,质谱分析其分子质量为422.99m/z。结论:从土样中分离得到粘细菌Myxococcus stipitatus STXZ77,从该菌中分离得到抗肿瘤活性组分AP-C,具有开发成抗肿瘤药物的潜在价值。
[Abstract]:Objective: to isolate and purify myxobacteria from soil samples, identify and classify myxobacteria in order to enrich the resources of myxobacteria and study their antitumor activity. Methods: inactivated Escherichia coli induction method was used to isolate and purify myxobacteria from soil samples and observe their morphology. Physiological and biochemical characteristics and homology analysis of 16s rRNA gene sequence were used to identify the strain. In vitro antitumor activity was studied by adding XAD-16 macroporous adsorption resin to the supernatant of fermentation broth to extract the crude extract of fermentation product (CCK-8). RP-HPLC was used to isolate the antitumor active component LC-MS / MS to analyze its molecular weight. Results: STXZ77 strains were isolated and identified. The strain was named Myxococcus stipitatus STXZ77.The fermentation product of this strain, XAD-16 resin crude extract, was applied to murine melanoma cell B16. Many kinds of tumor cells, such as mouse breast cancer cell 4T 1, human liver cancer cell SMMC-7721, human cervical cancer cell he Laand human colon cancer cell SW480, have good cytotoxicity. The ICP 50 values of the cells exposed for 24 hours were 5.34 渭 g / ml, 13.50 渭 g / ml, 11.93 渭 g / ml, 28.70 渭 g / ml, respectively. 48.09 渭 g / ml, and the toxicity of HUVEC to human umbilical vein endothelial cells was 17.09 渭 g / ml, less than B16. 4Cytotoxicity of T1 and SMMC-7721. RP-HPLC was used to isolate anti-tumor active component AP-C. The molecular weight of the product was 422.99m / z. Conclusion: Myxococcus stipitatus STXZ77 was isolated from soil samples. The antitumor active component AP-C was isolated from the strain, which has the potential value of developing antitumor drugs.
【作者单位】：湖南师范大学生命科学学院微生物分子生物学国家重点实验室培育基地;
【基金】：国家“973”计划(2012CB722301) 国家“863”计划(2011AA10A203) 湖南省生物发育工程及新产品研发协同创新中心项目(20134486)资助项目
【分类号】：R96;R914
【正文快照】： *国家“973”计划(2012CB722301),国家“863”计划(2011AA10A203),湖南省生物发育工程及新产品研发协同创新中心项目(20134486)资助项目粘细菌(Myxobacteria)是一类杆状、可滑行、具有复杂多细胞行为的革兰氏阴性菌,能够形成多细胞的子实体和抗逆性的粘孢子。粘细菌属于变形杆

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