科微学术

微生物学报

珠江口沉积物来源链霉菌SCSIO 40020中bafilomycins的鉴定及其生物合成基因簇的异源表达
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

广东省基础与应用基础研究基金(2021A1515010839);广州市科技计划(202102020513);国家自然科学基金(21907098,41706181);国家重点研发计划(2019YFC0312503);王宽诚教育基金(GJTD-2020-12)


Identification of bafilomycins and heterologous expression of its biosynthetic gene cluster from the Pearl River Estuary sediment-derived Streptomyces sp. SCSIO 40020
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    [目的] 从珠江口沉积物来源的菌株SCSIO 40020中分离bafilomycins,并对其生物合成基因簇进行克隆和异源表达研究。[方法] 通过分析菌株SCSIO 40020的16S rRNA基因序列并构建系统发育树以鉴定菌种,以柱层析法和制备色谱法对次级代谢产物进行分离纯化,借助波谱学手段完成单体化合物的结构鉴定,采用生物信息学分析定位bafilomycins的生物合成基因簇,通过筛选菌株SCSIO 40020基因组的细菌人工染色体文库和接合转移将bafilomycins生物合成基因簇导入3种链霉菌进行异源表达,利用高效液相色谱检测异源表达菌株的发酵产物。[结果] 菌株SCSIO 40020被鉴定为链霉菌属菌株,从其发酵产物中分离鉴定了2个单体化合物bafilomycins A1和D。克隆了链霉菌SCSIO 40020中bafilomycins的生物合成基因簇并推导了其生物合成途径,在3种链霉菌中表达产生了bafilomycins。[结论] 从珠江口环境中获得了一株产生bafilomycins的链霉菌SCSIO 40020,成功建立了该菌株次级代谢产物生物合成基因簇的异源表达体系,并首次在链霉菌Streptomyces lividans SBT18、Streptomyces coelicolor M1154和Streptomyces albus J1074中进行了表达,获得了bafilomycins,为后续bafilomycins结构类似物的生产和链霉菌SCSIO 40020中新结构活性化合物的挖掘奠定了基础。

    Abstract:

    [Objective] To identify the macrolide antibiotic bafilomycins from the strain SCSIO 40020 isolated from the Pearl River Estuary sediment. To clone and heterologously express the biosynthetic gene cluster (BGC) of bafilomycins.[Methods] The strain SCSIO 40020 was identified based on phylogenetic analysis of its 16S rRNA gene sequence. The generated compounds from SCSIO 40020 were purified via normal phase-column chromatography and semi-preparative chromatography. The chemical structures of the isolated compounds were subsequently elucidated by comprehensive spectroscopic analyses. The bafilomycins BGC was identified by using bioinformatics approach. Through screening the bacterial artificial chromosome (BAC) library of this strain, the BAC containing the bafilomycins BGC was introduced into three Streptomyces hosts by conjugation for expression. Afterwards, the fermentation culture of the recombinant strains were analyzed on high-performance liquid chromatography (HPLC). [Results] The strain SCSIO 40020 was identified as Streptomyces genus and two macrolide compounds bafilomycins A1and D were isolated from the fermentation crude extract. The bafilomycins BGC was cloned and successfully expressed in three heterologous Streptomyces hosts. In addition, the biosynthetic pathway of bafilomycins was proposed. [Conclusion] The bafilomycin-producing strain Streptomyces sp. SCSIO 40020 was obtained from the Pearl River Estuary. The BAC heterologous expression system of this strain was successfully established. Moreover, the bafilomycins BGC was successfully expressed in Streptomyces lividans SBT18, Streptomyces coelicolor M1154 and Streptomyces albus J1074 for the first time, which laid solid foundation for the production of bafilomycin analogues and mining other interesting BGCs from Streptomyces sp. SCSIO 40020 in the further study.

    参考文献
    相似文献
    引证文献
引用本文

翟诗岚,刘威,吴正超,朱义广,张文军,张长生,马亮.珠江口沉积物来源链霉菌SCSIO 40020中bafilomycins的鉴定及其生物合成基因簇的异源表达.微生物学报,2021,61(12):3991-4005

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2021-02-25
  • 最后修改日期:2021-04-28
  • 录用日期:
  • 在线发布日期: 2021-12-17
  • 出版日期: