Strain improvement of recombinant Escherichia coli for efficient production of plant flavonoids
Strain improvement of recombinant Escherichia coli for efficient production of plant flavonoids
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DOI：
10.1021/mp7001472
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发表时间：
2008-03-01
影响因子：
4.9
通讯作者：
Koffas, Mattheos A. G.
中科院分区：
文献类型：
--
作者：
Leonard, Effendi;Yan, Yajun;Koffas, Mattheos A. G.
植物类黄酮多酚继续发现越来越多的药物和营养品应用;然而，它们的分离，特别是从植物材料中分离纯化合物，仍然是一个潜在的挑战。在过去，大肠杆菌，最充分表征的微生物之一，已被用作蛋白质表达和小分子的异源生物合成的重组宿主。然而，在许多情况下，表达的蛋白质活性和生物合成效率受到宿主细胞特性的极大限制，例如前体和辅因子的可用性以及蛋白质或产物耐受性。在本工作中，我们开发了E。大肠杆菌菌株，通过传统的代谢工程技术能够高水平合成黄酮类化合物。除了嫁接植物生物合成途径之外，这些方法还包括替代碳同化途径的工程化和竞争性反应途径的抑制，以增加细胞内类黄酮骨架前体和辅因子。利用这种策略，我们报道了从苯丙酸和黄烷-3-的前体中分别产生高达700 mg/L的植物特异性黄烷酮和高达113 mg/L的花青素。这些结果证明了从E.用于制药和保健品应用。
Plant flavonoid polyphenols continue to find increasing pharmaceutical and nutraceutical applications; however their isolation, especially of pure compounds, from plant material remains an underlying challenge. In the past Escherichia coli, one of the most well-characterized microorganisms, has been utilized as a recombinant host for protein expression and heterologous biosynthesis of small molecules. However, in many cases the expressed protein activities and biosynthetic efficiency are greatly limited by the host cellular properties, such as precursor and cofactor availability and protein or product tolerance. In the present work, we developed E. coli strains capable of high-level flavonoid synthesis through traditional metabolic engineering techniques. In addition to grafting the plant biosynthetic pathways, the methods included engineering of an alternative carbon assimilation pathway and the inhibition of competitive reaction pathways in order to increase intracellular flavonoid backbone precursors and cofactors. With this strategy, we report the production of plant-specific flavanones up to 700 mg/L and anthocyanins up to 113 mg/L from phenylpropanoic acid and flavan-3-of precursors, respectively. These results demonstrated the efficient and scalable production of plant flavonoids from E. coli for pharmaceutical and nutraceutical applications.