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利用模块化代谢策略从葡萄糖中生产(2S)-松属素的大肠杆菌代谢工程。

Metabolic engineering of Escherichia coli for (2S)-pinocembrin production from glucose by a modular metabolic strategy.

机构信息

Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.

出版信息

Metab Eng. 2013 Mar;16:48-55. doi: 10.1016/j.ymben.2012.11.009. Epub 2012 Dec 14.

Abstract

Flavonoids are valuable natural products widely used in human health and nutrition. Recent advances in synthetic biology and metabolic engineering have yielded improved strain titers and yields. However, current fermentation strategies often require supplementation of expensive phenylpropanoic precursors in the media and separate evaluation of each strategy in turn as part of the flavonoid pathway, implicitly assuming the modifications are additive. In this study, an Escherichia coli fermentation system was developed to bypass both of these problems. An eight-step pathway, consisting of 3-deoxy-D-arabinoheptulosonate-7-phosphate synthase (DAHPS), chorismate mutase/prephenate dehydratase (CM/PDT), phenylalanine ammonia lyase (PAL), 4-coumarate:CoA ligase (4CL), chalcone synthase (CHS), chalcone isomerase (CHI), malonate synthetase, and malonate carrier protein, was assembled on four vectors in order to produce the flavonoid precursor (2S)-pinocembrin directly from glucose. Furthermore, a modular metabolic strategy was employed to identify conditions that optimally balance the four pathway modules. Once this metabolic balance was achieved, such strains were capable of producing 40.02mg/L (2S)-pinocembrin directly from glucose. These results were attained by culturing engineered cells in minimal medium without additional precursor supplementation. The fermentation platform described here paves the way for the development of an economical process for microbial production of flavonoids directly from glucose.

摘要

类黄酮是一种有价值的天然产物,广泛应用于人类健康和营养领域。近年来,合成生物学和代谢工程的发展提高了菌株的效价和产量。然而,目前的发酵策略通常需要在培养基中补充昂贵的苯丙氨酸前体,并依次单独评估每种策略,作为类黄酮途径的一部分,这隐含地假设这些修饰是累加的。在本研究中,开发了一种大肠杆菌发酵系统来解决这两个问题。该系统构建了一个由 8 步途径组成的途径,包括 3-脱氧-D-阿拉伯庚酮糖-7-磷酸合酶(DAHPS)、分支酸变位酶/预苯酸脱水酶(CM/PDT)、苯丙氨酸解氨酶(PAL)、4-香豆酸:CoA 连接酶(4CL)、查尔酮合酶(CHS)、查尔酮异构酶(CHI)、丙二酸合成酶和丙二酸载体蛋白,该途径组装在四个载体上,目的是直接从葡萄糖生产类黄酮前体(2S)-松属素。此外,还采用模块化代谢策略来确定最佳平衡四个途径模块的条件。一旦达到这种代谢平衡,这些菌株就能够直接从葡萄糖产生 40.02mg/L 的(2S)-松属素。这些结果是通过在没有额外前体补充的最小培养基中培养工程细胞获得的。这里描述的发酵平台为从葡萄糖直接微生物生产类黄酮开发经济的生产工艺铺平了道路。

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