期刊
MICROORGANISMS
卷 10, 期 3, 页码 -出版社
MDPI
DOI: 10.3390/microorganisms10030628
关键词
hyperoside; quercetin; UDP-dependent glycosyltransferase; UDP-glucose; metabolic engineering
类别
资金
- High-level talent project of West Anhui University [WGKQ2021025]
- National Natural Science Foundation of China [31901383]
- Natural Science Foundation of Anhui Province [2008085QB96]
- China Postdoctoral Science Foundation [2020M680134]
- Guangdong Basic and Applied Basic Research Foundation [2019A1515111051]
Hyperoside, a compound with higher bioactivities than quercetin, was efficiently synthesized from quercetin by co-expressing UGT from Petunia hybrida and GalE in E. coli. Additional enzymes Pgm and GalU were introduced to enhance UDP-glucose synthesis, leading to improved hyperoside production. Through fermentation optimization, hyperoside production was increased, and substrate-fed batch fermentation further increased production to 831.6 mg/L with a high conversion ratio and specific productivity. This efficient pathway can be applied for glycosylation of other flavonoids and bioactive substances.
Hyperoside (quercetin 3-O-galactoside) exhibits many biological functions, along with higher bioactivities than quercetin. In this study, three UDP-dependent glycosyltransferases (UGTs) were screened for efficient hyperoside synthesis from quercetin. The highest hyperoside production of 58.5 mg center dot L-1 was obtained in a recombinant Escherichia coli co-expressing UGT from Petunia hybrida (PhUGT) and UDP-glucose epimerase (GalE, a key enzyme catalyzing the conversion of UDP-glucose to UDP-galactose) from E. coli. When additional enzymes (phosphoglucomutase (Pgm) and UDP-glucose pyrophosphorylase (GalU)) were introduced into the recombinant E. coli, the increased flux toward UDP-glucose synthesis led to enhanced UDP-galactose-derived hyperoside synthesis. The efficiency of the recombinant strain was further improved by increasing the copy number of the PhUGT, which is a limiting step in the bioconversion. Through the optimization of the fermentation conditions, the production of hyperoside increased from 245.6 to 411.2 mg center dot L-1. The production was also conducted using a substrate-fed batch fermentation, and the maximal hyperoside production was 831.6 mg center dot L-1, with a molar conversion ratio of 90.2% and a specific productivity of 27.7 mg center dot L-1 center dot h(-1) after 30 h of fermentation. The efficient hyperoside synthesis pathway described here can be used widely for the glycosylation of other flavonoids and bioactive substances.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据