Efficient Synthesis of Eriodictyol from l-Tyrosine in Escherichia coli
Efficient Synthesis of Eriodictyol from l-Tyrosine in Escherichia coli
复制标题
大肠杆菌中 L-酪氨酸高效合成圣草酚
DOI：
10.1128/aem.03986-13
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发表时间：
2014-03
影响因子：
4.4
通讯作者：
Chen Jian
中科院分区：
文献类型：
--
作者：
Zhu Saijie;Wu Junjun;Du Guocheng;Zhou Jingwen;Chen Jian
摘要类黄酮类化合物对人类健康的益处日益引起人们的关注。由于从植物中提取高纯度的类黄酮是一个主要障碍，因此人们对利用微生物宿主进行生物合成产生了兴趣。Eriodictyol是一种具有抗炎和抗氧化活性的黄酮类化合物。它的有效合成受到两个因素的阻碍：细胞色素P450在大肠杆菌中的低表达和胞内丙二酰辅酶A(丙二酰辅酶A)的低浓度。为了解决这些问题，在大肠杆菌中表达了截短的植物P450类黄酮3‘-羟基酶(TF3’H)与截短的P450还原酶(TCPR)的融合蛋白。该融合蛋白可与酪氨酸解氨酶(TAL)、4-香豆酸-辅酶A连接酶(4CL)、查尔酮合成酶(CHS)和查尔酮异构酶(CHI)同时表达。此外，利用代谢工程来提高丙二酰辅酶A的利用率，以达到新的代谢平衡，并重新平衡基因的相对表达，以促进淫羊藿醇的积累。该方法使淫羊藿醇的产量比对照菌株高203%。在此基础上，L酪氨酸发酵生产淫羊藿醇的产量达到107 mg/L。本工作为在大肠杆菌中从L酪氨酸甚至葡萄糖中高效合成其他羟基黄酮类化合物提供了一种方法。
ABSTRACT The health benefits of flavonoids for humans are increasingly attracting attention. Because the extraction of high-purity flavonoids from plants presents a major obstacle, interest has emerged in biosynthesizing them using microbial hosts. Eriodictyol is a flavonoid with anti-inflammatory and antioxidant activities. Its efficient synthesis has been hampered by two factors: the poor expression of cytochrome P450 and the low intracellular malonyl coenzyme A (malonyl-CoA) concentration in Escherichia coli. To address these issues, a truncated plant P450 flavonoid, flavonoid 3′-hydroxylase (tF3′H), was functionally expressed as a fusion protein with a truncated P450 reductase (tCPR) in E. coli. This allowed the engineered E. coli to produce eriodictyol from l-tyrosine by simultaneously coexpressing the fusion protein with tyrosine ammonia lyase (TAL), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), and chalcone isomerase (CHI). In addition, metabolic engineering was employed to enhance the availability of malonyl-CoA so as to achieve a new metabolic balance and rebalance the relative expression of genes to enhance eriodictyol accumulation. This approach made the production of eriodictyol 203% higher than that in the control strain. By using these strategies, the production of eriodictyol from l-tyrosine reached 107 mg/liter. The present work offers an approach to the efficient synthesis of other hydroxylated flavonoids from l-tyrosine or even glucose in E. coli.
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铃木 - 米洛拉反应在类黄酮的合成中的应用。
DOI：
10.3390/molecules18044739
发表时间：
2013-04-22
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Selepe MA;Van Heerden FR
通讯作者：
Van Heerden FR
影响因子：
4.9
作者：
Cheng, Xuefang;Liu, Fang;Hao, Haiping
通讯作者：
Hao, Haiping
影响因子：
8.4
作者：
Santos, Christine Nicole S.;Koffas, Mattheos;Stephanopoulos, Gregory
通讯作者：
Stephanopoulos, Gregory
DOI：
10.1271/bbb.110952
发表时间：
2012-07
期刊：
Bioscience, Biotechnology, and Biochemistry
影响因子：
--
作者：
J. Yoo;Ji-Hee Kim;Sae-Jin Park;Yeo-Jin Kang;Tack-Joong Kim
通讯作者：
J. Yoo;Ji-Hee Kim;Sae-Jin Park;Yeo-Jin Kang;Tack-Joong Kim
影响因子：
8.4
作者：
Jones, Kristala L.;Kim, Seon-Won;Keasling, J. D.
通讯作者：
Keasling, J. D.