| 孙倩楠,张会鹏,侯路宽,等.深海链霉菌SCSIO 1071中聚酮合酶基因簇Cluster 46编码产物探究[J].中国海洋药物,2022,41(3):25-33. |
| 深海链霉菌SCSIO 1071中聚酮合酶基因簇Cluster 46编码产物探究 |
| The exploration of secondary metabolites of polyketide synthase gene cluster 46 from SCSIO 1071 |
| 投稿时间：2021-05-17 修订日期：2021-06-04 |
| DOI： |
| 中文关键词: 深海链霉菌 Streptomyces olivaceus SCSIO 1071 聚酮合酶生物合成基因簇 多重耐药菌 |
| English Keywords:deep-sea derived Streptomyces Streptomyces olivaceus SCSIO 1071 polyketide synthase biosynthesis gene cluster multiple drug resistant bacteria |
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| 中文摘要: |
| 目的 对南海深海来源链霉菌Streptomyces olivaceus SCSIO 1071中新颖I型聚酮合酶基因簇Cluster 46及其编码产物进行探究。方法 通过生物信息学手段对基因簇Cluster 46的基因组成进行分析；采用PCR-targeting策略构建聚酮合酶基因orf27和LuxR家族转录调控基因orf46双交换阻断突变株；通过高效液相色谱法(high performance liquid chromatography，HPLC)分析野生株和突变株发酵产物的差异,检测粗提物对3株多重耐药(multiple drug resistant, MDR)菌(Staphylococcus aureus CCARM 3090、Enterococcus faecalis CCARM 5172、Enterococcus faecium CCARM 5203)的抑菌活性。结果 得到了Δorf27和Δorf46双交换阻断突变株；HPLC分析结果表明,相比较野生株,Δorf27和Δorf46突变株中化合物峰1-10不再产生；突变株发酵液粗提物对3株MDR菌抑菌活性较野生株明显降低；液相色谱-质谱（liquid chromatograph mass spectrometer，LC-MS）数据和紫外（ultraviolet，UV）数据表明化合物峰1-10可能为dixiamycins类化合物。结论 orf27和orf46的阻断间接影响了具有抗MDR菌活性的化合物峰1-10的产生,为挖掘S. olivaceus SCSIO 1071中新颖活性次级代谢产物奠定了基础。 |
| English Summary: |
| Abstract: Objective To explore the secondary metabolites of novel type I polyketide synthase gene cluster 46 in deep-sea-derived Streptomyces olivaceus SCSIO 1071. Methods The gene organization of Cluster 46 was predicted by bioinformatics analysis. The polyketide synthase gene orf27 and the LuxR family regulator gene orf46 were knocked out by using PCR-targeting strategy. The fermentation broths were analyzed by high performance liquid chromatography (HPLC). In addition, the crude extracts were subjected to bioassay against three multi-drug resistance (MDR) bacteria strains (Staphylococcus aureus CCARM 3090, Enterococcus faecalis CCARM 5172, Enterococcus faecium CCARM 5203). Results HPLC analysis suggested that the production of compounds 1-10 were abolished in Δorf27 and Δorf46 mutant strains. The crude extracts from mutant strains exhibited much lower inhibition activities against three MDR bacteria strains. Liquid chromatograph mass spectrometer (LC-MS) and ultraviolet (UV) analysis suggested that compounds 1-10 might be dixiamycins. Conclusion Blocking of orf27 and orf46 indirectly affected the production of compounds 1-10 with anti-MDR bacteria activities, laying a foundation for the discovery of new active secondary metabolites from S. olivaceus SCSIO 1071. |
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