铁载体
生物合成
链霉菌
化学
电喷雾电离
天然产物
铜
非核糖体肽
基因簇
生物化学
突变体
质谱法
生物
色谱法
基因
细菌
有机化学
遗传学
作者
Lijuan Wang,Menglin Zhu,Qingbo Zhang,Xu Zhang,Panlei Yang,Zihui Liu,Yun Deng,Yiguang Zhu,Xueshi Huang,Li Han,Shengqing Li,Jialin He
标识
DOI:10.1021/acschembio.7b00897
摘要
A nonribosomal peptide synthetase (NRPS) gene cluster (sfa) was identified in Streptomyces thioluteus to direct the biosynthesis of the diisonitrile antibiotic SF2768. Its biosynthetic pathway was reasonably proposed based on bioinformatics analysis, metabolic profiles of mutants, and the elucidation of the intermediate and shunt product structures. Bioinformatics-based alignment found a putative ATP-binding cassette (ABC) transporter related to iron import within the biosynthetic gene cluster, which implied that the product might be a siderophore. However, characterization of the metal-binding properties by high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), metal–ligand titration, thin-layer chromatography (TLC), and chrome azurol S (CAS) assays revealed that the final product SF2768 and its diisonitrile derivatives specifically bind copper, rather than iron, to form stable complexes. Inductively coupled plasma mass spectrometry (ICP-MS) analysis revealed that the intracellular cupric content of S. thioluteus significantly increased upon incubation with the copper–SF2768 complex, direct evidence for the copper acquisition function of SF2768. Further in vivo functional characterization of the transport elements for the copper–SF2768 complexes not only confirmed the chalkophore identity of the compound but also gave initial clues into the copper uptake mechanism of this nonmethanotrophic microorganism.
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