Chimeric Leader Peptides for the Generation of Non-Natural Hybrid RiPP Products.
Chimeric Leader Peptides for the Generation of Non-Natural Hybrid RiPP Products.
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DOI：
10.1021/acscentsci.7b00141
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发表时间：
2017-06-28
影响因子：
18.2
通讯作者：
Mitchell DA
中科院分区：
文献类型：
--
作者：
Burkhart BJ;Kakkar N;Hudson GA;van der Donk WA;Mitchell DA
组合来自多个途径的生物合成酶是产生具有所需结构特征的分子的有吸引力的方法;然而，进展受到来自不相关途径的酶的不相容性和对替代底物的不耐受性的阻碍。核糖体合成和后修饰的肽（RiPP）是一种多样的天然产物类别，其采用高度适合于工程化新化合物的生物合成逻辑。RiPP生物合成蛋白通过与通常位于前体肽的N-末端前导区的基序结合来修饰其底物。在这里，我们利用这一特点，通过设计前导肽，使识别和处理的多个酶从不相关的RiPP途径。使用这种广泛适用的策略，噻唑啉形成环己基转移酶与来自sactipeptide和lanthipeptide家族的酶组合，以产生新的天然杂合RIPP。我们还提供了深入的设计功能，使控制杂交生物合成，优化酶的相容性，并建立一个通用的平台，工程额外的杂交RIPP。我们报告了一种通用的方法，使酶的组合从不同的RiPP途径的合理设计的新的混合postperturbationally修饰的肽。
Combining biosynthetic enzymes from multiple pathways is an attractive approach for producing molecules with desired structural features; however, progress has been hampered by the incompatibility of enzymes from unrelated pathways and intolerance toward alternative substrates. Ribosomally synthesized and posttranslationally modified peptides (RiPPs) are a diverse natural product class that employs a biosynthetic logic that is highly amenable to engineering new compounds. RiPP biosynthetic proteins modify their substrates by binding to a motif typically located in the N-terminal leader region of the precursor peptide. Here, we exploit this feature by designing leader peptides that enable recognition and processing by multiple enzymes from unrelated RiPP pathways. Using this broadly applicable strategy, a thiazoline-forming cyclodehydratase was combined with enzymes from the sactipeptide and lanthipeptide families to create new-to-nature hybrid RiPPs. We also provide insight into design features that enable control over the hybrid biosynthesis to optimize enzyme compatibility and establish a general platform for engineering additional hybrid RiPPs. We report a general approach that enables combination of enzymes from different RiPP pathways for the rational design of new hybrid posttranslationally modified peptides.