Biosynthetic Gene Cluster of a D-Tryptophan-Containing Lasso Peptide, MS-271
Biosynthetic Gene Cluster of a D-Tryptophan-Containing Lasso Peptide, MS-271
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DOI：
10.1002/cbic.201800315
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发表时间：
2018-10-04
期刊：
影响因子：
3.2
通讯作者：
Dairi, Tohru
中科院分区：
文献类型：
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作者：
Feng, Zhi;Ogasawara, Yasushi;Dairi, Tohru
MS-271由链霉菌M-271产生，是一种套索肽天然产物，含有21个氨基酸残基，在其C末端具有D-色氨酸。由于套索肽是核糖体肽，MS-271的生物合成，特别是D-Trp的引入机制，是非常感兴趣的。MS-271生物合成基因簇通过MS-271生产者的基因组测序草图鉴定，并且揭示了前体肽包含包括C-末端色氨酸的所有21个氨基酸残基。这表明D-Trp残基通过差向异构化引入。在前体肽基因的侧翼区发现了修饰酶的基因，例如大环内酰胺合成酶（mslC）、前体肽识别元件（mslB 1）、半胱氨酸蛋白酶（mslB 2）、二硫氧化还原酶（mslE、mslF）和未知功能的蛋白质（mslH）。虽然明显的差向异构酶基因在集群中不存在，异源表达的推定MS-271集群中变铅青链霉菌表明，它包含所有必要的基因MS-271生产，包括一个新的肽差向异构酶的基因。此外，基因缺失实验表明，MS-271的生产中，MslB 1，-B2，-C和-H是必不可少的，一些生物合成酶的相互作用是必不可少的MS-271的生物合成。
MS-271, produced by Streptomyces sp. M-271, is a lasso peptide natural product comprising 21 amino acid residues with a D-tryptophan at its C terminus. Because lasso peptides are ribosomal peptides, the biosynthesis of MS-271, especially the mechanism of D-Trp introduction, is of great interest. The MS-271 biosynthetic gene cluster was identified by draft genome sequencing of the MS-271 producer, and it was revealed that the precursor peptide contains all 21 amino acid residues including the C-terminal tryptophan. This suggested that the D-Trp residue is introduced by epimerization. Genes for modification enzymes such as a macrolactam synthetase (mslC), precursor peptide recognition element (mslB1), cysteine protease (mslB2), disulfide oxidoreductases (mslE, mslF), and a protein of unknown function (mslH) were found in the flanking region of the precursor peptide gene. Although obvious epimerase genes were absent in the cluster, heterologous expression of the putative MS-271 cluster in Streptomyces lividans showed that it contains all the necessary genes for MS-271 production including a gene for a new peptide epimerase. Furthermore, a gene-deletion experiment indicated that MslB1, -B2, -C and -H were indispensable for MS-271 production and that some interactions of the biosynthetic enzymes were essential for the biosynthesis of MS-271.