| 陈霖锋,程永猛,肖菲,等.深海冷泉链霉菌Streptomyces albus OUCLQ4-3中albonoursin类似物的分离鉴定及生物合成途径研究[J].中国海洋药物,2025,(5):-. |
| 深海冷泉链霉菌Streptomyces albus OUCLQ4-3中albonoursin类似物的分离鉴定及生物合成途径研究 |
| Identification of albonoursin derivatives from cold-seep-derived Streptomyces albus OUCLQ4-3 and their biosynthetic pathways |
| 投稿时间：2024-05-09 修订日期：2024-07-03 |
| DOI： |
| 中文关键词: 深海冷泉链霉菌 次级代谢产物 抗多重耐药菌活性 环二肽合酶生物合成途径 |
| English Keywords:deep-sea cold-seep derived Streptomyces secondary metabolites anti multi-drug-resistant (MDR) bacterial activity CDPS biosynthetic pathways |
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| 中文摘要: |
| 目的 探究深海冷泉来源微生物产生次级代谢产物的潜力,挖掘具有抗多重耐药(multi-drug resistant, MDR)菌活性的次级代谢产物,并研究其生物合成途径。方法 采用OSMAC策略结合抗菌活性测试确定发酵培养基,采用有机溶剂萃取、C18反相硅胶开放柱层析、半制备高效液相(high-performance liquid chromatography,HPLC)等分离手段对发酵产物进行分离纯化,通过质谱(mass spectrum,MS)、核磁(Nuclear magnetic resonance,NMR)数据以及文献比对进行化合物的结构鉴定,并对其进行抗MDR菌活性测试；对菌株进行全基因组测序,定位活性化合物生物合成基因簇,采用生物信息学分析和异源表达手段探究化合物的生物合成途径。结果 从深海冷泉链霉菌Streptomyces albus OUCLQ4-3发酵产物中分离得到2个albonoursin类似物1-N-methyl-(E,Z)-albonoursin(1)和lansai D(2)；活性结果显示,化合物1和2对Klebsiella pneumoniae ATCC 13883具有抑制活性；定位了化合物1和2的生物合成基因簇,通过异源表达证明了环二肽合酶Orf3的功能,推测了化合物1和2的生物合成途径。结论 从深海冷泉链霉菌S. albus OUCLQ4-3发酵产物中分离鉴定了2个具有优良抗多重耐药菌活性的albonoursin类似物1和2；基于生物信息学分析和异源表达推测了1和2的生物合成是由环二肽合酶Orf3催化形成cyclo(L-Phe-L-Leu)(cFL)骨架、环二肽氧化酶Orf1和Orf2催化脱氢形成双键、甲基转移酶Orf4催化N-甲基化完成。 |
| English Summary: |
| Objective To explore the potential of deep-sea cold-seep derived microorganisms to produce secondary metabolites, and explore secondary metabolites with anti multidrug-resistant (MDR) bacterial activity, and study their biosynthetic pathways. Methods Different culture media were used for fermentation, the culture medium with the best antibacterial activity was selected for large-scale fermentation. The fermentation extract was separated by solvent extraction, C18 reverse phase silica gel chromatography and semi-preparative high performance liquid chromatography (HPLC). The compounds structures were identified by spectroscopic analysis (NMR and MS) as well as comparison with published researches, followed by anti-MDR activities tests. The strain was subjected to whole genome sequencing, the active compound biosynthesis gene cluster was located, and the biosynthesis pathways of the compounds were explored using bioinformatics analysis and heterologous expression methods. Results Two albonoursin analogues, 1-N-methyl-(E,Z)-albonoursin (1) and lansai D (2) were isolated from the fermentation products of Streptomyces albus OUCLQ4-3 in deep-sea cold-seep; The activity results showed that compounds 1 and 2 exhibited inhibitory activity against Klebsiella pneumoniae ATCC 13883; The biosynthetic gene clusters of compounds 1 and 2 were located, and the function of the cyclic dipeptide synthase Orf3 was demonstrated through heterologous expression, suggesting the biosynthetic pathways of compounds 1 and 2. Conclusion Compounds 1 and 2 with excellent anti multidrug-resistant bacterial activity were isolated from the fermentation products of Streptomyces albus OUCLQ4-3.Based on bioinformatics analysis and heterologous expression, it is speculated that the biosynthesis of compounds 1 and 2 are catalyzed by the cyclodipeptide synthase Orf3 to form the cyclo(L-Phe-L Leu)(cFL) skeleton, the cyclodipeptide oxidase Orf1 and Orf2 catalyze dehydrogenation to form double bonds, and the methyltransferase Orf4 catalyzes N-methylation. Conclusion S. albus OUCLQ4-3 was screened from deep-sea cold-seep, and three compounds with excellent anti multidrug-resistant activity were isolated and identified from their fermentation products. Based on bioinformatics analysis and heterologous expression, it is speculated that the biosynthetic pathways of diketopiperazine compounds 1 and 2 are catalyzed by one molecule of L-leucine tRNA and one molecule of L-phenylalanine tRNA through cyclic dipeptide synthase Orf3, cyclic dipeptide oxidase Orf1 and Orf2, and methyltransferase Orf4 to form compound 1 and compound 2. |
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