Heterologous production of new lasso peptide koreensin based on genome mining
Heterologous production of new lasso peptide koreensin based on genome mining
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DOI：
10.1038/s41429-020-00363-5
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发表时间：
2020-08-27
影响因子：
3.3
通讯作者：
Kodani, Shinya
中科院分区：
文献类型：
--
作者：
Fuwa, Hiroki;Hemmi, Hikaru;Kodani, Shinya
套索肽是一类核糖体生物合成和翻译后修饰的肽，以结结构为共同基序。通过对韩国鞘单胞菌基因组的分析，发现了一个新的拉索肽合成基因簇。有趣的是，前体肽基因的氨基酸序列包括两个细胞粘附基序序列（KGD和DGR）。利用s的隐性生物合成基因簇进行了新的套索肽的异源生产。koreensis。结果表明，该基因表达系统在地下鞘氨单胞菌中产生了一种新的拉索肽，命名为koreenin。通过核磁共振和质谱分析确定了koreensin的结构。基于NOE实验和耦合常数得到了koreensin的三维结构。通过定点诱变实验，采用异种生产方法制备了具有RGD而非KGD的koreensin-RGD。Koreensin和Koreensin - rgd虽然具有细胞粘附基序，但没有表现出细胞粘附抑制活性。在koreenin的基序之间可能存在盐桥，它可能阻止细胞粘附基序的功能。
Lasso peptides are a class of ribosomally biosynthesized and posttranslationally modified peptides with a knot structure as a common motif. Based on a genome search, a new biosynthetic gene cluster of lasso peptide was found in the genome of the proteobacteriumSphingomonas koreensis. Interestingly, the amino acid sequence of the precursor peptide gene includes two cell adhesion motif sequences (KGD and DGR). Heterologous production of the new lasso peptide was performed using the cryptic biosynthetic gene cluster ofS. koreensis. As a result, a new lasso peptide named koreensin was produced by the gene expression system in the host strainSphingomonas subterranea. The structure of koreensin was determined by NMR and ESI-MS analysis. The three-dimensional structure of koreensin was obtained based on an NOE experiment and the coupling constants. A variant peptide (koreensin-RGD), which had RGD instead of KGD, was produced by heterologous production with site-directed mutagenesis experiment. Koreensin and koreensin-RGD did not show cell adhesion inhibitory activity, although the molecules possessed cell adhesion motifs. The possible presence of a salt bridge between the motifs in koreensin was indicated, and it may prevent the cell adhesion motif from functioning.