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首页> 外文期刊>Applied Microbiology and Biotechnology >Metabolic engineering of Escherichia coli for the biosynthesis of flavonoid-O-glucuronides and flavonoid-O-galactoside
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Metabolic engineering of Escherichia coli for the biosynthesis of flavonoid-O-glucuronides and flavonoid-O-galactoside

机译：大黄素-O-葡糖醛酸苷和类黄酮-O-半乳糖苷生物合成的大肠杆菌代谢工程

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摘要

Most flavonoids are glycosylated and the nature of the attached sugar can strongly affect their physiological properties. Although many flavonoid glycosides have been synthesized in Escherichia coli, most of them are glucosylated. In order to synthesize flavonoids attached to alternate sugars such as glucuronic acid and galactoside, E. coli was genetically modified to express a uridine diphosphate (UDP)-dependent glycosyltransferase (UGT) specific for UDP-glucuronic acid (AmUGT10 from Antirrhinum majus or VvUGT from Vitis vinifera) and UDP-galactoside (PhUGT from Petunia hybrid) along with the appropriate nucleotide biosynthetic genes to enable simultaneous production of their substrates, UDP-glucuronic acid and UDP-galactose. To engineer UDP-glucuronic acid biosynthesis, the araA gene encoding UDP-4-deoxy-4-formamido-L-arabinose formyltransferase/UDP-glucuronic acid C-4aEuro(3) decarboxylase, which also used UDP-glucuronic acid as a substrate, was deleted in E. coli, and UDP-glucose dehydrogenase (ugd) gene was overexpressed to increase biosynthesis of UDP-glucuronic acid. Using these strategies, luteolin-7-O-glucuronide and quercetin-3-O-glucuronide were biosynthesized to levels of 300 and 687 mg/L, respectively. For the synthesis of quercetin 3-O-galactoside, UGE (encoding UDP-glucose epimerase from Oryza sativa) was overexpressed along with a glycosyltransferase specific for quercetin and UDP-galactose. Using this approach, quercetin 3-O-galactoside was successfully synthesized to a level of 280 mg/L.
机译：大多数类黄酮均被糖基化，附着糖的性质会强烈影响其生理特性。尽管在大肠杆菌中已经合成了许多类黄酮苷，但是它们中的大多数都是糖基化的。为了合成与替代糖（例如葡萄糖醛酸和半乳糖苷）相连的类黄酮，对大肠杆菌进行了基因修饰，以表达对尿苷二磷酸酯（UDP）依赖性的糖基转移酶（UGT），该酶对UDP-葡萄糖醛酸（Antirrhinum majus的AmUGT10或VvUGT葡萄）和UDP-半乳糖苷（矮牵牛杂种的PhUGT）以及适当的核苷酸生物合成基因，可同时生产其底物，UDP-葡萄糖醛酸和UDP-半乳糖。为了工程化UDP-葡萄糖醛酸的生物合成，编码UDP-4-脱氧-4-甲酰胺基-L-阿拉伯糖甲酰基转移酶/ UDP-葡萄糖醛酸C-4aEuro（3）脱羧酶的araA基因，也使用UDP-葡萄糖醛酸作为底物，在大肠杆菌中缺失了该基因，并且UDP-葡萄糖脱氢酶（ugd）基因被过表达以增加UDP-葡萄糖醛酸的生物合成。使用这些策略，生物合成木犀草素-7-O-葡糖醛酸和槲皮素-3-O-葡糖醛酸的水平分别为300和687 mg / L。为了合成槲皮素3-O-半乳糖苷，UGE（编码来自稻米的UDP-葡萄糖差向异构酶）与槲皮素和UDP-半乳糖特异的糖基转移酶一起过表达。使用这种方法，槲皮素3-O-半乳糖苷已成功合成至280 mg / L的水平。

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