Shao Zengyi, Rao Guodong, Li Chun, Abil Zhanar, Luo Yunzi, Zhao Huimin
Department of Chemical and Biomolecular Engineering, ‡Department of Chemistry, and §Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States.
ACS Synth Biol. 2013 Nov 15;2(11):662-9. doi: 10.1021/sb400058n. Epub 2013 Aug 22.
Natural products (secondary metabolites) are a rich source of compounds with important biological activities. Eliciting pathway expression is always challenging but extremely important in natural product discovery because an individual pathway is tightly controlled through a unique regulation mechanism and hence often remains silent under the routine culturing conditions. To overcome the drawbacks of the traditional approaches that lack general applicability, we developed a simple synthetic biology approach that decouples pathway expression from complex native regulations. Briefly, the entire silent biosynthetic pathway is refactored using a plug-and-play scaffold and a set of heterologous promoters that are functional in a heterologous host under the target culturing condition. Using this strategy, we successfully awakened the silent spectinabilin pathway from Streptomyces orinoci. This strategy bypasses the traditional laborious processes to elicit pathway expression and represents a new platform for discovering novel natural products.
天然产物（次生代谢物）是具有重要生物活性的化合物的丰富来源。在天然产物发现中，引发途径表达一直具有挑战性但极其重要，因为单个途径通过独特的调控机制受到严格控制，因此在常规培养条件下通常保持沉默。为了克服传统方法缺乏普遍适用性的缺点，我们开发了一种简单的合成生物学方法，该方法将途径表达与复杂的天然调控解耦。简而言之，使用即插即用支架和一组在目标培养条件下在异源宿主中起作用的异源启动子对整个沉默的生物合成途径进行重构。利用这一策略，我们成功唤醒了来自橄榄链霉菌的沉默的spectinabilin途径。该策略绕过了引发途径表达的传统繁琐过程，代表了一个发现新型天然产物的新平台。
