Isolation and structural characterization of capistruin, a lasso peptide predicted from the genome sequence of Burkholderia thailandensis E264
Isolation and structural characterization of capistruin, a lasso peptide predicted from the genome sequence of Burkholderia thailandensis E264
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DOI：
10.1021/ja802966g
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发表时间：
2008-08-27
影响因子：
15
通讯作者：
Marahiel, Mohamed A.
中科院分区：
文献类型：
--
作者：
Knappe, Thomas A.;Linne, Uwe;Marahiel, Mohamed A.
Lasso肽是一类结构独特的生物活性肽，其特征在于打结排列，其中C-末端穿过N-末端大环内酰胺环。虽然核糖体合成，只有基因簇的最好的研究套索肽MccJ 25从大肠杆菌组成的前体蛋白McjA和加工和免疫蛋白McjB，McjC，和McjD是已知的。通过基因组挖掘研究，我们已经确定了所有四个蛋白质在伯克霍尔德菌E264的同源物，并预测该菌株产生套索肽。在这里，我们报告的预测肽，命名capistruin的成功分离。在优化发酵条件后，质谱和NMR结构研究证明capistruin采用新的套索折叠。在大肠杆菌中的套索肽的异源生产表明，所鉴定的基因足以用于capistruin的生物合成，capistruin对密切相关的伯克霍尔德氏菌和假单胞菌菌株表现出抗微生物活性。总的来说，我们的合理方法应该广泛适用于新的套索肽的分离，以探索其高结构稳定性和多样的生物活性。
Lasso peptides are a structurally unique class of bioactive peptides characterized by a knotted arrangement, where the C-terminus threads through an N-terminal macrolactam ring. Although ribosomally synthesized, only the gene cluster for the best studied lasso peptide MccJ25 from Escherichia coli consisting of the precursor protein McjA and the processing and immunity proteins McjB, McjC, and McjD is known. Through genome mining studies, we have identified homologues of all four proteins in Burkholderia thailandensis E264 and predicted this strain to produce a lasso peptide. Here we report the successful isolation of the predicted peptide, named capistruin. Upon optimization of the fermentation conditions, mass spectrometric and NMR structural studies proved capistruin to adopt a novel lasso fold. Heterologous production of the lasso peptide in Escherichia coli showed that the identified genes are sufficient for the biosynthesis of capistruin, which exhibits antimicrobial activity against closely related Burkholderia and Pseudomonas strains. In general, our rational approach should be widely applicable for the isolation of new lasso peptides to explore their high structural stability and diverse biological activity.