Department of Chemistry, North Carolina State University, Raleigh, NC 27695-8204, USA.
Org Biomol Chem. 2013 Jul 21;11(27):4449-58. doi: 10.1039/c3ob40633d.
Combinatorial biosynthesis approaches that involve modular type I polyketide synthases (PKSs) are proven strategies for the synthesis of polyketides. In general however, such strategies are usually limited in scope and utility due to the restricted substrate specificity of polyketide biosynthetic machinery. Herein, a panel of chemo-enzymatically synthesized acyl-CoA's was used to probe the promiscuity of a polyketide synthase. Promiscuity determinants were dissected, revealing that the KS is remarkably tolerant to a diverse array of extender units, while the AT likely discriminates between extender units that are native to the producing organism. Our data provides a clear blueprint for future enzyme engineering efforts, and sets the stage for harnessing extender unit promiscuity by employing various in vivo polyketide diversification strategies.
组合生物合成方法涉及模块化的 I 型聚酮合酶（PKS），是聚酮类化合物合成的有效策略。然而，通常情况下，由于聚酮生物合成机制的底物特异性受到限制，此类策略的应用范围和实用性受到限制。在此，我们使用一组化学酶合成的酰基辅酶 A 来研究聚酮合酶的混杂性。对混杂性决定因素进行了剖析，结果表明 KS 对各种扩展单元具有显著的耐受性，而 AT 可能会区分产生菌中原有的扩展单元。我们的数据为未来的酶工程努力提供了明确的蓝图，并为利用各种体内聚酮多样化策略来利用扩展单元的混杂性奠定了基础。
