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喙尾琵琶甲肠道来源链霉菌(Streptomyces sp.)BPA71次级代谢产物的分离鉴定及其生物活性评价
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云南省基础研究项目(202001AU070022);云南省中青年学术和技术带头人后备人才项目(202305AC160035,202105AC160062);云南省重大科技专项计划(202002AA100007)


Isolation, identification, and bioactivity evaluation of secondary metabolites from Blaps rynchopetera gut-derived Streptomyces sp. BPA71
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    摘要:

    【背景】昆虫是世界上种类最多、肠道菌群资源最丰富且多样的动物类群之一。昆虫肠道微生物具有产生活性次级代谢产物的能力,是活性天然产物的重要来源。【目的】研究药用昆虫喙尾琵琶甲(Blaps rynchopetera)成虫肠道来源链霉菌(Streptomyces sp.) BPA71的次级代谢产物及其生物活性。【方法】利用正相硅胶柱色谱、葡聚糖凝胶Sephadex LH-20柱色谱等方法分离纯化该菌株的发酵粗提物,采用牛津杯法进行抗菌活性追踪,确定抗菌活性部位,通过ESI-MS、NMR等波谱数据分析对化合物结构进行鉴定,采用微量肉汤稀释法测定最低抑菌浓度(minimal inhibitory concentration, MIC),采用MTS法测定抗肿瘤活性。【结果】从Streptomyces sp. BPA71的固体发酵提取物中共分离得到4个已知化合物,通过对比核磁数据确定为糠酸甲酯(1)、吡咯甲酰胺A (2)、吡咯甲酰胺B (3)和吲哚-3-乙酸甲酯(4)。抗菌活性结果显示化合物2具有广谱抗菌活性。此外,化合物2对宫颈癌细胞HeLa、肺癌细胞A549、肝癌细胞SMMC-7721、乳腺癌细胞MDA-MB-231和结肠癌细胞SW480这5株肿瘤细胞均有明显的抑制活性。【结论】喙尾琵琶甲肠道来源Streptomyces sp. BPA71可产生丰富的生物活性物质,该研究结果为进一步挖掘喙尾琵琶甲肠道链霉菌的活性天然产物奠定了基础,同时丰富了人们对喙尾琵琶甲肠道微生物的认识。

    Abstract:

    [Background] Insects are the animal group with the most species and the most abundant gut microbiota resources in the world. The microorganisms in insect gut have the ability to produce active secondary metabolites, which is an important source of active natural products.[Objective] To identify the secondary metabolites produced by Streptomyces sp. BPA71 isolated from the gut of the medicinal insect Blaps rynchopetera and determine their biological activities. [Methods] The crude fermentation extract of the strain was separated and purified by column chromatography with silica gel and Sephadex LH-20. Oxford cup method was employed to determine the antimicrobial activity, and the antimicrobial active part was determined. The compound structure was identified by comparison of data from ESI-MS, 1H NMR, and 13C NMR. The minimum inhibitory concentration (MIC) and antitumor activity were determined by microbroth dilution method and MTS method, respectively.[Results] Four known compounds were purified from the crude fermentation extract of BPA71 and identified as methyl 2-furoate (1), pyrroloformamide A (2), pyrroloformamide B (3), and methyl 2-(1H-indol-3-yl) acetate (4). Compound 2 had broad-spectrum antimicrobial activity. In addition, it demonstrated significant inhibitory activities on five tumor cell lines, including cervical cancer cell line HeLa, lung cancer cell line A549, liver cancer cell line SMMC-7721, breast cancer cell line MDA-MB-231, and colon cancer cell line SW480. [Conclusion] Streptomyces sp. BPA71 from the gut of B. rynchopetera can produce rich bioactive substances. The findings of this study lay a foundation for further mining the active natural products of Streptomyces from the gut of B. rynchopetera and enriched our understanding of the gut microbiota of B. rynchopetera.

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李铭晖,王明明,殷鹏凯,肖怀,杨大松,杨银河. 喙尾琵琶甲肠道来源链霉菌(Streptomyces sp.)BPA71次级代谢产物的分离鉴定及其生物活性评价[J]. 微生物学通报, 2023, 50(5): 2076-2086

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  • 收稿日期:2023-01-06
  • 最后修改日期:
  • 录用日期:2023-03-10
  • 在线发布日期: 2023-05-06
  • 出版日期: 2023-05-20
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