Phylogeny-guided genome mining of roseocin family lantibiotics to generate improved variants of roseocin.
Phylogeny-guided genome mining of roseocin family lantibiotics to generate improved variants of roseocin.
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玫瑰菌素家族脂肪生物的系统发育引导的基因组挖掘可产生改善的玫瑰霉素变体。
DOI：
10.1186/s13568-023-01536-9
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发表时间：
2023-03-20
期刊：
影响因子：
3.7
通讯作者：
Sareen, Dipti
中科院分区：
文献类型：
--
作者：
Chaudhary, Sandeep;Kishen, Shweta;Singh, Mangal;Jassal, Sunanda;Pathania, Reeva;Bisht, Kalpana;Sareen, Dipti
Roseocin是来自玫瑰孢链霉菌（Streptomycesroseosporus）的双肽羊毛硫抗生素，在肽中携带广泛的分子内（甲基）羊毛硫氨酸桥连，并且对临床相关的革兰氏阳性病原体表现出协同抗菌活性。两种肽都具有保守的前导序列，但具有不同的核心区域。玫瑰霉素的生物合成涉及通过单一混杂羊毛硫肽合成酶RosM对两种前体肽进行翻译后修饰，以在Rosα核心中安装不可或缺的二硫键沿着分别在Rosα和Rosβ核心中安装四个和六个硫醚环。在放线菌门中的RosM同源物在这里被鉴定，以揭示玫瑰霉素家族的另外12个成员，其分化成三种类型的生物合成基因簇（BGC）。此外，BGC变体之间的进化速率和核心肽与前导肽内的变异性分析揭示了门依赖性羊毛硫肽进化。水平基因转移的分析揭示了其在核心肽多样性产生中的作用。仔细比对从开采的新型BGC中鉴定的玫瑰霉素肽的天然存在的不同同源物，以鉴定核心肽区域中的保守位点和取代。对Rosα肽中的这些选择位点进行突变以允许取代，在E. coli，并在体内进行RosM后修饰。尽管产生的变体数量有限，但与野生型玫瑰霉素相比，两种变体Rosα L 8 F和Rosα L 8 W以物种依赖性方式表现出显著改善的抑制活性。我们的研究证明，自然界中存在玫瑰菌素进化变体的天然储存库，并且关键变体可用于产生改进的变体。在线版本包含补充材料，可通过10.1186/s13568-023-01536-9获得。分类学上遥远的物种具有玫瑰菌素家族羊毛硫抗生素的广泛多样性。生物学引导的方法为RIPP生物工程开辟了新的可能性。使用半体外重构产生玫瑰霉素的生物活性变体。在线版本包含补充材料，可通过10.1186/s13568-023-01536-9获得。
Roseocin, the two-peptide lantibiotic from Streptomyces roseosporus, carries extensive intramolecular (methyl)lanthionine bridging in the peptides and exhibits synergistic antibacterial activity against clinically relevant Gram-positive pathogens. Both peptides have a conserved leader but a diverse core region. The biosynthesis of roseocin involves post-translational modification of the two precursor peptides by a single promiscuous lanthipeptide synthetase, RosM, to install an indispensable disulfide bond in the Rosα core along with four and six thioether rings in Rosα and Rosβ cores, respectively. RosM homologs in the phylum actinobacteria were identified here to reveal twelve other members of the roseocin family which diverged into three types of biosynthetic gene clusters (BGCs). Further, the evolutionary rate among the BGC variants and analysis of variability within the core peptide versus leader peptide revealed a phylum-dependent lanthipeptide evolution. Analysis of horizontal gene transfer revealed its role in the generation of core peptide diversity. The naturally occurring diverse congeners of roseocin peptides identified from the mined novel BGCs were carefully aligned to identify the conserved sites and the substitutions in the core peptide region. These selected sites in the Rosα peptide were mutated for permitted substitutions, expressed heterologously in E. coli, and post-translationally modified by RosM in vivo. Despite a limited number of generated variants, two variants, RosαL8F and RosαL8W exhibited significantly improved inhibitory activity in a species-dependent manner compared to the wild-type roseocin. Our study proves that a natural repository of evolved variants of roseocin is present in nature and the key variations can be used to generate improved variants. The online version contains supplementary material available at 10.1186/s13568-023-01536-9. Taxonomically distant species have a wide diversity of roseocin family lantibiotics. The phylogeny-guided approach opens new possibilities for RiPPs bioengineering. Bioactive variants of roseocin were generated using semi-in vitro reconstitution. The online version contains supplementary material available at 10.1186/s13568-023-01536-9.
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影响因子：
3.8
作者：
Barbosa, Joana;Caetano, Tania;Mendo, Sonia
通讯作者：
Mendo, Sonia
影响因子：
18.2
作者：
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作者：
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作者：
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5.2
作者：
Field D;Cotter PD;Hill C;Ross RP
通讯作者：
Ross RP