Bravo-Rodriguez Kenny, Klopries Stephan, Koopmans Kyra R M, Sundermann Uschi, Yahiaoui Samir, Arens Julia, Kushnir Susanna, Schulz Frank, Sanchez-Garcia Elsa
Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470 Mülheim an der Ruhr, Germany.
Fakultät für Chemie und Biochemie, Organische Chemie 1, Ruhr-Universität Bochum, Universitätsstraße 150, 44780 Bochum, Germany.
Chem Biol. 2015 Nov 19;22(11):1425-1430. doi: 10.1016/j.chembiol.2015.02.008. Epub 2015 Oct 29.
Polyketides are natural products frequently used for the treatment of various diseases, but their structural complexity hinders efficient derivatization. In this context, we recently introduced enzyme-directed mutasynthesis to incorporate non-native extender units into the biosynthesis of erythromycin. Modeling and mutagenesis studies led to the discovery of a variant of an acyltransferase domain in the erythromycin polyketide synthase capable of accepting a propargylated substrate. Here, we extend molecular rationalization of enzyme-substrate interactions through modeling, to investigate the incorporation of substrates with different degrees of saturation of the malonic acid side chain. This allowed the engineered biosynthesis of new erythromycin derivatives and the introduction of additional mutations into the AT domain for a further shift of the enzyme's substrate scope. Our approach yields non-native polyketide structures with functional groups that will simplify future derivatization approaches, and provides a blueprint for the engineering of AT domains to achieve efficient polyketide synthase diversification.
聚酮化合物是常用于治疗各种疾病的天然产物，但其结构复杂性阻碍了有效的衍生化。在此背景下，我们最近引入了酶导向的突变合成法，将非天然延伸单元纳入红霉素的生物合成中。建模和诱变研究导致在红霉素聚酮合酶中发现了一种能够接受炔丙基化底物的酰基转移酶结构域变体。在这里，我们通过建模扩展了酶-底物相互作用的分子合理化，以研究丙二酸侧链不同饱和度底物的掺入情况。这使得新红霉素衍生物的工程生物合成成为可能，并在酰基转移酶结构域中引入额外的突变，以进一步扩大酶的底物范围。我们的方法产生了具有官能团的非天然聚酮化合物结构，这将简化未来的衍生化方法，并为工程改造酰基转移酶结构域以实现有效的聚酮合酶多样化提供了蓝图。
