Modularity of RiPP Enzymes Enables Designed Synthesis of Decorated Peptides.
Modularity of RiPP Enzymes Enables Designed Synthesis of Decorated Peptides.
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DOI：
10.1016/j.chembiol.2015.06.014
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发表时间：
2015-07-23
影响因子：
--
通讯作者：
Schmidt EW
中科院分区：
文献类型：
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作者：
Sardar D;Lin Z;Schmidt EW
大环化酶和其他翻译后酶为制药和生物技术应用提供了具有改进性质的衍生肽。在这里，我们问是否可以同时控制和匹配多个翻译后修饰，以合理地产生新的肽衍生物。我们重建的cyanobactin肽天然产物在体外与多达五种不同的翻译后酶在一个单一的管。通过操纵酶的添加顺序和身份并利用其广泛的底物耐受性，我们设计了高度非天然衍生物的生产，包括长度为22个氨基酸的N-C肽大环。除了工程，这项工作更好地定义了大环化机制，提供了丝氨酸/苏氨酸翻译后异戊烯化的第一个生化证明，是第一个例子重建一个天然的，多步骤的RiPP途径与多个酶在一锅。总之，这项工作表明如何翻译后修饰酶的模块化可以用来设计和合成所需的肽基序。
Macrocyclases and other posttranslational enzymes afford derived peptides with improved properties for pharmaceutical and biotechnological applications. Here, we asked whether multiple posttranslational modifications could be simultaneously controlled and matched to rationally generate new peptide derivatives. We reconstituted the cyanobactin peptide natural products in vitro with up to five different posttranslational enzymes in a single tube. By manipulating the order of addition and identity of enzymes and exploiting their broad-substrate tolerance, we engineered the production of highly unnatural derivatives, including an N-C peptide macrocycle of 22 amino acids in length. In addition to engineering, this work better defines the macrocyclization mechanism, provides the first biochemical demonstration of Ser/Thr posttranslational prenylation, and is the first example of reconstitution of a native, multistep RiPP pathway with multiple enzymes in one pot. Overall, this work demonstrates how the modularity of posttranslational modification enzymes can be used to design and synthesize desirable peptide motifs.