Enhancing the modularity of the modular polyketide synthases:: Transacylation in modular polyketide synthases catalyzed by malonyl-CoA:ACP transacylase
Enhancing the modularity of the modular polyketide synthases:: Transacylation in modular polyketide synthases catalyzed by malonyl-CoA:ACP transacylase
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DOI：
10.1021/ja037429l
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发表时间：
2003-11-26
影响因子：
15
通讯作者：
Khoslaa, C
中科院分区：
文献类型：
--
作者：
Kumar, P;Koppisch, AT;Khoslaa, C
模块化聚酮合酶中延伸单元的选择性掺入主要由酰基转移酶 (AT) 结构域控制。 AT 结构域催化同一模块中从酰基辅酶 A 延伸单元到酰基载体蛋白 (ACP) 结构域的磷酸泛硫氨酸臂的转酰基作用。可以在工程模块中以最小的结构或动力学扰动调节单个模块的延伸单元特异性的新方法对于新型天然产物类似物的有效生物合成是可取的。我们已经证明，通过丙二酰辅酶A：ACP转酰基酶将丙二酰基团转酰基到突变型模块化聚酮化合物合酶的AT无效形式上，是用于位点特异性修饰的聚酮化合物的工程生物合成的有效策略。利用这种策略，6-脱氧赤酮内酯 B 合酶被设计为专门生产 2-去甲基-6-脱氧赤酮内酯 B。修饰后的系统在体外和体内的生产力与野生型合酶相当。
Selective incorporation of extender units in modular polyketide synthases is primarily controlled by acyl transferase (AT) domains. The AT domains catalyze transacylation of the extender unit from acyl-CoA to the phosphopantetheine arm of an acyl carrier protein (ACP) domain in the same module. New methods that can modulate the extender unit specificity of individual modules with minimal structural or kinetic perturbations in the engineered module are desirable for the efficient biosynthesis of novel natural product analogues. We have demonstrated that transacylation of malonyl groups onto an AT-null form of a mutant modular polyketide synthase by malonyl-CoA:ACP transacylase is an effective strategy for the engineered biosynthesis of site specifically modified polyketides. Using this strategy, 6-deoxyerythronolide B synthase was engineered to exclusively produce 2-desmethyl-6-deoxyerythronolide B. The productivity of the modified system was comparable to that of the wild-type synthase in vitro and in vivo.