于东琪,侯路宽,李花月,等.深海链霉菌Streptomyces somaliensis SCSIO ZH66 ptm生物合成基因簇中orf7功能的探究[J].中国海洋药物,2021,40(3):15-21. |
深海链霉菌Streptomyces somaliensis SCSIO ZH66 ptm生物合成基因簇中orf7功能的探究 |
Function of orf7 in the ptm biosynthetic gene cluster from the deep-sea-derived Strptomyces somaliensis SCSIO ZH66 |
投稿时间:2020-05-28 修订日期:2020-06-01 |
DOI: |
中文关键词: 深海链霉菌 多环特特拉姆酸大环内酰胺 天然产物 |
English Keywords:deepsea-derived Streptomyces polycyclic tetramate macrolactams natural products |
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中文摘要: |
目的 探究深海链霉菌Streptomyces somaliensis SCSIO ZH66中编码多环特特拉姆酸大环内酰胺(polycyclic tetramate macrolactams,PTMs)类化合物somamycin A-E的生物合成基因簇ptm中orf7对该基因簇编码产物的影响。方法 采用PCR-targeting策略将ptm基因簇中的orf7进行阻断,然后将野生株与突变株在相同条件下发酵后通过HPLC分析比较代谢产物的差异。 结果 生物信息学分析结果表明orf7的编码产物为细胞色素P450,实验成功得到了阻断突变株Δorf7, 通过发酵产物HPLC分析发现,orf7被阻断后,somamycin D的产量提高,而其14位羰基氧化物somamycin C则不再产生。 结论 说明ptm基因簇中细胞色素P450基因orf7在PTMs生物合成过程中参与了14位碳上的氧化修饰。 |
English Summary: |
Objective To explore the in vivo function of the putative cytochrome P450 gene orf7 in the polycyclic tetramate macrolactam biosynthetic gene cluster ptm encoding somamycins, from the deepsea-derived Streptomyces somaliensis SCSIO ZH66. Methods orf7 was inactivated by using PCR-targeting strategy. The mutant and the wild-type strains were fermented under the same conditions, and the fermentation broths were analyzed by HPLC. Results Bioinformatic analysis showed orf7 encodes a putative cytochrome P450. orf7 was knocked out successfully. HPLC analysis suggested that the disruption of orf7 led to yield increase of somamycin D, while its C14 oxide somamycin C no longer produced. Conclusion orf7 is probably involved in oxidization of the C14 hydroxyl group of somamycin D to carbonyl group. |
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