Heterologous expression of a cryptic gene cluster from Marinomonas fungiae affords a novel tricyclic peptide marinomonasin
Heterologous expression of a cryptic gene cluster from Marinomonas fungiae affords a novel tricyclic peptide marinomonasin
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DOI：
10.1007/s00253-021-11545-y
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发表时间：
2021-09-04
影响因子：
5
通讯作者：
Kodani, Shinya
中科院分区：
文献类型：
--
作者：
Kaweewan, Issara;Nakagawa, Hiroyuki;Kodani, Shinya
含ω-酯肽（OEPs）是一类核糖体合成和后修饰的肽（RIPPs）。含ω-酯肽的生物合成基因簇通常包括ATP-抓取连接酶编码基因，并且分布在各种细菌的基因组中。在海洋变形杆菌Marinomonasfungiae基因组序列中发现了一个新的含ω-酯肽生物合成基因簇。利用大肠杆菌表达宿主菌BL 21（DE 3）中的生物合成基因簇，实现了一种新的三环肽（marinomonasin）的异源生产。通过ESI-MS和NMR实验，确定了marinomonasin的结构是一个18个氨基酸长的三环肽，分子中有一个酯键和两个异肽键。通过对HMBC和NOESY数据的解释，确定了三种分子内键的桥接模式。在含ω-酯的肽组中，marinomonasin的桥接模式是前所未有的。结果表明，用于生产海单胞菌素的ATP-抓取连接酶是一种新型酶，具有形成一个酯键和两个异肽键的双功能活性。
The omega-ester-containing peptides (OEPs) are a group of ribosomally synthesized and post-translationally modified peptides (RiPPs). The biosynthetic gene clusters of omega-ester-containing peptides commonly include ATP-grasp ligase coding genes and are distributed over the genomes of a wide variety of bacteria. A new biosynthetic gene cluster of omega-ester-containing peptides was found in the genome sequence of the marine proteobacterium Marinomonas fungiae. Heterologous production of a new tricyclic peptide named marinomonasin was accomplished using the biosynthetic gene cluster in Escherichia coli expression host strain BL21(DE3). By ESI-MS and NMR experiments, the structure of marinomonasin was determined to be a tricyclic peptide 18 amino acids in length with one ester and two isopeptide bonds in the molecule. The bridging patterns of the three intramolecular bonds were determined by the interpretation of HMBC and NOESY data. The bridging pattern of marinomonasin was unprecedented in the omega-ester-containing peptide group. The results indicated that the ATP-grasp ligase for the production of marinomonasin was a novel enzyme possessing bifunctional activity to form one ester and two isopeptide bonds.