| 侯婧,刘晶,刘增智,等.多效调控基因wblAyo对海洋链霉菌Streptomyces youssoufiensis OUC6819次级代谢产物及形态分化的影响[J].中国海洋药物,2018,37(5):13-20. |
| 多效调控基因wblAyo对海洋链霉菌Streptomyces youssoufiensis OUC6819次级代谢产物及形态分化的影响 |
| Effect of the global regulator wblAyo on secondary metabolites and morphological differentiation of Streptomyces youssoufiensis OUC6819 |
| 投稿时间：2018-05-25 修订日期：2018-06-22 |
| DOI： |
| 中文关键词: wblAyo 次级代谢产物 形态分化 多重耐药菌 |
| English Keywords:wblAyo secondary metabolites morphological differentiation MDR strains |
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| 中文摘要: |
| 目的 激活南海红树林来源的链霉菌Streptomyces youssoufiensis OUC6819中不表达或表达水平低的隐性基因簇，进一步挖掘新颖的次级代谢产物。方法 采用PCR-targeting策略敲除WhiB-like（Wbl）家族的wblAyo基因，通过HPLC分析野生株和突变株发酵产物的差异，检测发酵液粗提物和C18开放柱层析组分对5种多重耐药（MDR）菌（Staphylococcus aureus CCARM 3090, Salmonella typhimurium CCARM 8250, Escherichia coli CCARM 1009, Enterococcus faecium CCARM 5203，Enterococcus faecalis CCARM 5172）的抑制活性，通过固体平板实验和光学显微镜观察链霉菌形态分化的改变。结果 得到了正确的ΔwblAyo阻断突变株，HPLC分析结果表明：与野生株相比，突变株中化合物峰1和2分别提高了15.6和9.3倍；突变株发酵液粗提物和组分11对E. faecium CCARM 5203，E. faecalis CCARM 5172和S. aureus CCARM 3090的抗菌活性与野生株相比明显提高，而且突变株丧失了产生孢子的能力。结论 wblAyo参与了S. youssoufiensis OUC6819的次级代谢和形态分化过程。wblAyo的阻断激活了S. youssoufiensis OUC6819中抗MDR菌活性次级代谢产物的产生，为继续挖掘活性化合物提供了必要基础，还为其他海洋链霉菌中隐性基因簇的激活提供了参考。 |
| English Summary: |
| Objective To activate cryptic biosynthetic gene clusters in mangrove-derived Streptomyces youssoufiensis OUC6819, and thus to discover new secondary metabolites. Methods The Wbl family regulator wblAyo was knocked out by PCR-targeting strategy. The mutant and wild-type strains were fermented under the same conditions, and the fermentation broths were then analyzed by HPLC. In addition, the antibacterial activities of the crude extracts and fractions (Fr) were tested against five multi-drug resistance (MDR) strains (S. aureus CCARM 3090, S. typhimurium CCARM 8250, E. coli CCARM 1009, E. faecium CCARM 5203 and E. faecalis CCARM 5172). The morphological differentiation of the strains was observed by using phase contrast microscope. Result HPLC analysis suggested that the yields of compound peaks 1 and 2 were increased by about 15.6- and 9.3-fold, respectively, in the ΔwblAyo strain compared to those in the wild-type strain. The crude extracts and Fr 11 of the mutant exhibited much stronger inhibition against E. faecium CCARM 5203, E. faecalis CCARM 5172 and S. aureus CCARM 3090. In addition, the mutant was abolished sporulation. Conclusion wblAyo is involved in the regulation of secondary metabolism and morphological differentiation in S. youssoufiensis OUC6819, and the disruption of wblAyo activated the production of compounds with anti-MDR bacteria strains activity. These results offered a foundation for discovery of new bioactive compounds and a strategy for activating cryptic gene clusters in other marine Streptomyces stains. |
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