Steiniger Charlotte, Hoffmann Sylvester, Süssmuth Roderich D
Institut für organische Chemie , Technische Universität Berlin , Strasse des 17. Juni 124/TC2 , Berlin , 10623 , Germany.
ACS Synth Biol. 2019 Apr 19;8(4):661-667. doi: 10.1021/acssynbio.8b00541. Epub 2019 Mar 19.
Nonribosomal peptide synthetases assemble a considerable number of structurally complex peptides of pharmacological importance. This turns them into important biosynthetic machineries for peptide diversification by engineering approaches. To date, manifold reprogramming approaches focus on employing module and domain exchanges, or the engineering of domains responsible for amino acid recognition. In this work, we present an engineering strategy for the manipulation of product assembly modes by fusing iterative fungal cyclodepsipeptide synthetases. The reassignment of terminal condensation domains as canonical condensation domains induces a switch from an exclusively iterative into a mixed linear/iterative peptide assembly mode. In the heterologous host E. coli we thus produced in vivo novel hybrid cyclodepsipeptides with altered structural symmetry. Our findings contribute a new experimental set of nonribosomal peptide synthetase reprogramming to the engineering toolbox for peptide structure diversification.
非核糖体肽合成酶可组装大量具有药理学重要性的结构复杂的肽。这使其通过工程方法成为肽多样化的重要生物合成机制。迄今为止，多种重新编程方法侧重于采用模块和结构域交换，或对负责氨基酸识别的结构域进行工程改造。在这项工作中，我们提出了一种通过融合迭代型真菌环缩肽合成酶来操纵产物组装模式的工程策略。将末端缩合结构域重新指定为典型缩合结构域会诱导从完全迭代的肽组装模式转变为混合线性/迭代的肽组装模式。因此，在异源宿主大肠杆菌中，我们在体内产生了具有改变的结构对称性的新型杂合环缩肽。我们的研究结果为肽结构多样化的工程工具箱贡献了一组新的非核糖体肽合成酶重新编程的实验方法。
