SBIR Phase I:Metabolic Engineering of Isoflavonoid Biosynthesis in Eschericia coli

这项小型企业创新研究第一阶段的研究将创建一种生物合成系统，用于经济、环境友好、多功能地生产天然和非天然异黄酮。通过将从植物中合成异黄酮类化合物的最佳酶转移到大肠杆菌中，这项研究将创造出自我复制的代谢机器，将廉价的原料转化为由所使用的前体和添加的酶确定的各种异黄酮类化合物中的任何一种。具体地说，这项研究将共同开发异黄酮类生物合成中的关键酶，并开发一种适应这些酶的特殊需求的大肠杆菌菌株。这将提供一个支架，可以在其上添加其他酶来同时创建新的异黄酮类化合物及其生产方法。异黄酮类化合物是豆类中的天然化合物，具有改善人类健康的巨大潜力。最简单的异黄酮类化合物，染料木素和大豆苷元，因其抗氧化和抗癌活性而引起临床关注。更复杂的异黄酮类化合物，如葛根素和芒柄花素，也正在被评估用于营养食品或制药应用。受到异黄酮类化合物改变人类细胞过程的多种方式的吸引，化学家们已经开始合成新一代非天然的、基于异黄酮类化合物的候选药物，这些候选药物可能比它们的天然前体更有效和更特异。事实上，其中一些已经在进行临床试验。异黄酮类化合物的供应是基于从植物中提取，这一过程既不有效也不一致，以及化学合成，这种合成对某些分子很好，但对其他分子来说效率低，例如那些在异黄酮核结构上特定位置含有糖或烷基的分子。这一过程将扩大这些化合物的可获得性，并为国家的利益提供新产品。

This Small Business Innovation Research Phase I research will create a biosynthetic system for economical, environmentally benign, versatile production of isoflavonoids, natural and non-natural. By transferring the optimal enzymes for the synthesis of isoflavonoids from plants into E. coli, the research will create self-replicating metabolic machines that convert inexpensive feedstocks into any of a diverse array of isoflavonoids determined by the precursors used and the enzymes added. Specifically, the research will co-develop the key enzymes in the biosynthesis of isoflavonoids and an E. coli strain adapted for the particular needs of these enzymes. This will provide a scaffold upon which other enzymes can be added to simultaneously create new isoflavonoids and the means of their production. Isoflavonoids are natural compounds from legumes that have great potential for improving human health. The simplest isoflavonoids , genistein and daidzein, have attracted clinical attention for their antioxidant and anticancer activities. More complex isoflavonoids such as puerarin and formononetin are also being evaluated for nutraceutical or pharmaceutical applications. Attracted by the numerous ways that isoflavonoids alter human cellular processes, chemists have begun to synthesize new generations of non-natural, isoflavonoid-based pharmaceutical candidates that might be more effective and specific than their natural precursors. In fact, some are already in clinical trials. The supply of isoflavonoids is based on extraction from plants, a process that is neither efficient nor consistent, and chemical synthesis which is excellent for some molecules but inefficient for others such as those that contains sugars or alkyl groups at specific positions on the isoflavonoid core structure. This process will expand the availability of these compounds and provide novel products for the benefit of the country.