Heterologous Production of Glidobactins/Luminmycins in Escherichia coli Nissle Containing the Glidobactin Biosynthetic Gene Cluster from Burkholderia DSM7029
Heterologous Production of Glidobactins/Luminmycins in Escherichia coli Nissle Containing the Glidobactin Biosynthetic Gene Cluster from Burkholderia DSM7029
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DOI：
10.1002/cbic.201402199
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发表时间：
2014-10-13
期刊：
影响因子：
3.2
通讯作者：
Zhang, Youming
中科院分区：
文献类型：
--
作者：
Bian, Xiaoying;Huang, Fan;Zhang, Youming
天然产物肽基蛋白酶体抑制剂作为抗癌药物具有很大的潜力。本研究从伯克霍尔德菌dsm7029中克隆了一种强效蛋白酶体抑制剂glidobactin的生物合成基因簇，并在大肠杆菌鼻中成功检测到glidobactin /luminmycins。我们还通过在异源宿主中改变启动子将glidobactin A的产量提高了10倍。此外，通过对比质谱/质谱裂解分析鉴定了两个新的生物合成中间体。无环发光霉素E的鉴定表明TE结构域对环化具有底物特异性。sybactins的异种表达体系的建立，为通过分子工程技术产生新的sybactins作为蛋白酶体抑制剂提供了基础，但TE结构域的特异性不容忽视。
Natural product peptide-based proteasome inhibitors show great potential as anticancer drugs. Here we have cloned the biosynthetic gene cluster of a potent proteasome inhibitorglidobactin from Burkholderia DSM7029and successfully detected glidobactins/luminmycins in E. coli Nissle. We have also improved the yield of glidobactin A tenfold by promoter change in a heterologous host. In addition, two new biosynthetic intermediates were identified by comparative MS/MS fragmentation analysis. Identification of acyclic luminmycin E implies substrate specificity of the TE domain for cyclization. The establishment of a heterologous expression system for syrbactins provided the basis for the generation of new syrbactins as proteasome inhibitors by molecular engineering, but the TE domain's specificity cannot be ignored.