Expanding the chemical space of synthetic cyclic peptides using a promiscuous macrocyclase from prenylagaramide biosynthesis
Expanding the chemical space of synthetic cyclic peptides using a promiscuous macrocyclase from prenylagaramide biosynthesis
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使用来自异戊二烯酰胺生物合成的混杂大环化酶扩展合成环肽的化学空间
DOI：
10.1101/2020.03.17.996157
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
E. Schmidt
中科院分区：
文献类型：
--
作者：
Snigdha Sarkar;W. Gu;E. Schmidt
环肽是极好的候选药物，将大环化反应置于药物开发的顶峰。来自蓝藻蛋白生物合成的PatG和相关的双作用蛋白酶负责切断c末端识别序列并将底物大环化以提供环状肽。该反应已被发现用于酶促合成各种大环。然而，这些酶在以非蛋白质源性噻唑/噻唑啉残基为末端的底物上发挥最佳作用，使合成策略复杂化。在这里，我们的生物化学特征的一类新的patg样大环化酶天然使用脯氨酸，避免了额外的化学或生化步骤的必要性。我们通过实验确定了合成广泛存在的类丙烯酰胺天然产物的生化步骤，包括大环化和丙烯酰化。利用饱和诱变，我们发现大环化酶PagG和戊烯基转移酶PagF是高度混杂的，在体外产生了100多个环肽及其戊烯基衍生物的文库。通过将我们的结果与已知的蓝藻蛋白大环酶进行比较，我们编目了一系列应该合成大多数小大环的酶。总的来说，这些数据表明，通过选择正确的蓝藻蛋白大环酶，可以获得大量的酶合成大环。
Cyclic peptides are excellent drug candidates, placing macrocyclization reactions at the apex of drug development. PatG and related dual-action proteases from cyanobactin biosynthesis are responsible for cleaving off the C-terminal recognition sequence and macrocyclizing the substrate to provide cyclic peptides. This reaction has found use in the enzymatic synthesis of diverse macrocycles. However, these enzymes function best on substrates that terminate with the non-proteinogenic thiazole/thiazoline residue, complicating synthetic strategies. Here, we biochemically characterize a new class of PatG-like macrocyclases that natively use proline, obviating the necessity of additional chemical or biochemical steps. We experimentally define the biochemical steps involved in synthesizing the widespread prenylagaramide-like natural products, including macrocyclization and prenylation. Using saturation mutagenesis, we show that macrocyclase PagG and prenyltransferase PagF are highly promiscuous, producing a library of more than 100 cyclic peptides and their prenylated derivatives in vitro. By comparing our results to known cyanobactin macrocyclase enzymes, we catalog a series of enzymes that collectively should synthesize most small macrocycles. Collectively, these data reveal that, by selecting the right cyanobactin macrocyclase, a large array of enzymatically synthesized macrocycles are accessible.
影响因子：
15
作者：
Sardar D;Hao Y;Lin Z;Morita M;Nair SK;Schmidt EW
通讯作者：
Schmidt EW
影响因子：
--
作者：
Gu W;Dong SH;Sarkar S;Nair SK;Schmidt EW
通讯作者：
Schmidt EW