Streamlined Chemoenzymatic Synthesis of Cyclic Peptides by Non-ribosomal Peptide Cyclases
Streamlined Chemoenzymatic Synthesis of Cyclic Peptides by Non-ribosomal Peptide Cyclases
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DOI：
10.1021/jacs.2c11082
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发表时间：
2023-01-13
影响因子：
15
通讯作者：
Wakimoto, Toshiyuki
中科院分区：
文献类型：
--
作者：
Kobayashi, Masakazu;Fujita, Kei;Wakimoto, Toshiyuki
大环化改善了多肽的药学性质；然而，区域环化和化学选择性分子内环化仍然具有挑战性。在这里，我们开发了一种简化的化学酶方法，通过利用非核糖体肽(NRP)环化酶来合成环肽。通过C-端二醇酯官能团连接到树脂上的线型多肽是在固体载体上合成的，以避免在液相中将基团安装到多肽底物上，这往往会引发不必要的差向异构化。裂解树脂结合的多肽可以得到纯度足够高的二醇酯，可以很容易地以头尾相接的方式环化，这是一种典型的青霉素结合蛋白型硫代酯酶(PBP-type TE)。对同源野生型酶的探索以及合理的蛋白质工程拓宽了酶促大内酰胺的范围。这种方法可能会加速NRP环酶作为生物催化剂的开发。
Macrocyclization improves the pharmaceutical properties of peptides; however, regio- and chemoselective intramolecular cyclizations remain challenging. Here we developed a streamlined chemoenzymatic approach to synthesize cyclic peptides by exploiting non-ribosomal peptide (NRP) cyclases. Linear peptides linked to the resin through a C-terminal diol ester functionality are synthesized on a solid support, to circumvent the installation of leaving groups to the peptidic substrates in the liquid phase which often triggers undesirable epimerization. Cleavage of the resin-bound peptides yielded the diol esters with sufficient purity to be readily cyclized in a head-to-tail manner by SurE, a representative penicillin-binding protein-type thioesterase (PBP-type TE). Explorations of homologous wild-type enzymes as well as rational protein engineering have broadened the scope of the enzymatic macrolactamization. This method will potentially accelerate the exploitation of NRP cyclases as biocatalysts.