利用大肠杆菌系统工程高效生产原儿茶酸。

IF 6.8 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic engineering Pub Date : 2024-02-16 DOI:10.1016/j.ymben.2024.02.003
Ming Wang , Haomiao Wang , Cong Gao , Wanqing Wei , Jia Liu , Xiulai Chen , Guipeng Hu , Wei Song , Jing Wu , Fan Zhang , Liming Liu
{"title":"利用大肠杆菌系统工程高效生产原儿茶酸。","authors":"Ming Wang ,&nbsp;Haomiao Wang ,&nbsp;Cong Gao ,&nbsp;Wanqing Wei ,&nbsp;Jia Liu ,&nbsp;Xiulai Chen ,&nbsp;Guipeng Hu ,&nbsp;Wei Song ,&nbsp;Jing Wu ,&nbsp;Fan Zhang ,&nbsp;Liming Liu","doi":"10.1016/j.ymben.2024.02.003","DOIUrl":null,"url":null,"abstract":"<div><p>Protocatechuic acid (3, 4-dihydroxybenzoic acid, PCA) is widely used in the pharmaceuticals, health food, and cosmetics industries owing to its diverse biological activities. However, the inhibition of 3-dehydroshikimate dehydratase (AroZ) by PCA and its toxicity to cells limit the efficient production of PCA in <em>Escherichia coli</em>. In this study, a high-level strain of 3-dehydroshikimate, <em>E. coli</em> DHS01, was developed by blocking the carbon flow from the shikimate-overproducing strain <em>E. coli</em> SA09. Additionally, the PCA biosynthetic pathway was established in DHS01 by introducing the high-activity <em>Ap</em>AroZ. Subsequently, the protein structure and catalytic mechanism of 3-dehydroshikimate dehydratase from <em>Acinetobacter pittii</em> PHEA-2 (<em>Ap</em>AroZ) were clarified. The variant <em>Ap</em>AroZ<sup>R363A</sup>, achieved by modulating the conformational dynamics of <em>Ap</em>AroZ, effectively relieved product inhibition. Additionally, the tolerance of the strain <em>E. coli</em> PCA04 to PCA was enhanced by adaptive laboratory evolution, and a biosensor-assisted high-throughput screening method was designed and implemented to expedite the identification of high-performance PCA-producing strains. Finally, in a 5 L bioreactor, the final strain PCA05 achieved the highest PCA titer of 46.65 g/L, a yield of 0.23 g/g, and a productivity of 1.46 g/L/h for PCA synthesis from glucose using normal fed-batch fermentation. The strategies described herein serve as valuable guidelines for the production of other high-value and toxic products.</p></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"82 ","pages":"Pages 134-146"},"PeriodicalIF":6.8000,"publicationDate":"2024-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Efficient production of protocatechuic acid using systems engineering of Escherichia coli\",\"authors\":\"Ming Wang ,&nbsp;Haomiao Wang ,&nbsp;Cong Gao ,&nbsp;Wanqing Wei ,&nbsp;Jia Liu ,&nbsp;Xiulai Chen ,&nbsp;Guipeng Hu ,&nbsp;Wei Song ,&nbsp;Jing Wu ,&nbsp;Fan Zhang ,&nbsp;Liming Liu\",\"doi\":\"10.1016/j.ymben.2024.02.003\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Protocatechuic acid (3, 4-dihydroxybenzoic acid, PCA) is widely used in the pharmaceuticals, health food, and cosmetics industries owing to its diverse biological activities. However, the inhibition of 3-dehydroshikimate dehydratase (AroZ) by PCA and its toxicity to cells limit the efficient production of PCA in <em>Escherichia coli</em>. In this study, a high-level strain of 3-dehydroshikimate, <em>E. coli</em> DHS01, was developed by blocking the carbon flow from the shikimate-overproducing strain <em>E. coli</em> SA09. Additionally, the PCA biosynthetic pathway was established in DHS01 by introducing the high-activity <em>Ap</em>AroZ. Subsequently, the protein structure and catalytic mechanism of 3-dehydroshikimate dehydratase from <em>Acinetobacter pittii</em> PHEA-2 (<em>Ap</em>AroZ) were clarified. The variant <em>Ap</em>AroZ<sup>R363A</sup>, achieved by modulating the conformational dynamics of <em>Ap</em>AroZ, effectively relieved product inhibition. Additionally, the tolerance of the strain <em>E. coli</em> PCA04 to PCA was enhanced by adaptive laboratory evolution, and a biosensor-assisted high-throughput screening method was designed and implemented to expedite the identification of high-performance PCA-producing strains. Finally, in a 5 L bioreactor, the final strain PCA05 achieved the highest PCA titer of 46.65 g/L, a yield of 0.23 g/g, and a productivity of 1.46 g/L/h for PCA synthesis from glucose using normal fed-batch fermentation. The strategies described herein serve as valuable guidelines for the production of other high-value and toxic products.</p></div>\",\"PeriodicalId\":18483,\"journal\":{\"name\":\"Metabolic engineering\",\"volume\":\"82 \",\"pages\":\"Pages 134-146\"},\"PeriodicalIF\":6.8000,\"publicationDate\":\"2024-02-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Metabolic engineering\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1096717624000193\",\"RegionNum\":1,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Metabolic engineering","FirstCategoryId":"5","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1096717624000193","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

原儿茶酸(3, 4-二羟基苯甲酸,PCA)具有多种生物活性,被广泛应用于制药、保健食品和化妆品行业。然而,PCA 对 3-脱氢莽草酸脱水酶(AroZ)的抑制作用及其对细胞的毒性限制了大肠杆菌生产 PCA 的效率。在本研究中,通过阻断莽草酸过量产生菌株大肠杆菌 SA09 的碳流,培育出了 3-脱氢莽草酸高产菌株大肠杆菌 DHS01。此外,通过引入高活性 ApAroZ,在 DHS01 中建立了 PCA 生物合成途径。随后,阐明了皮氏不动杆菌 PHEA-2 的 3-脱氢莽草酸脱水酶(ApAroZ)的蛋白质结构和催化机理。通过调节 ApAroZ 的构象动力学而获得的变体 ApAroZR363A 能有效缓解产物抑制。此外,通过实验室适应性进化,提高了大肠杆菌 PCA04 菌株对 PCA 的耐受性,并设计和实施了一种生物传感器辅助的高通量筛选方法,以加快鉴定高性能 PCA 生产菌株。最后,在一个 5 L 的生物反应器中,最终菌株 PCA05 通过正常的喂料批次发酵,从葡萄糖合成 PCA 的最高滴度为 46.65 g/L,产量为 0.23 g/g,生产率为 1.46 g/L/h。本文所述的策略可作为生产其他高价值和有毒产品的宝贵指南。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Efficient production of protocatechuic acid using systems engineering of Escherichia coli

Protocatechuic acid (3, 4-dihydroxybenzoic acid, PCA) is widely used in the pharmaceuticals, health food, and cosmetics industries owing to its diverse biological activities. However, the inhibition of 3-dehydroshikimate dehydratase (AroZ) by PCA and its toxicity to cells limit the efficient production of PCA in Escherichia coli. In this study, a high-level strain of 3-dehydroshikimate, E. coli DHS01, was developed by blocking the carbon flow from the shikimate-overproducing strain E. coli SA09. Additionally, the PCA biosynthetic pathway was established in DHS01 by introducing the high-activity ApAroZ. Subsequently, the protein structure and catalytic mechanism of 3-dehydroshikimate dehydratase from Acinetobacter pittii PHEA-2 (ApAroZ) were clarified. The variant ApAroZR363A, achieved by modulating the conformational dynamics of ApAroZ, effectively relieved product inhibition. Additionally, the tolerance of the strain E. coli PCA04 to PCA was enhanced by adaptive laboratory evolution, and a biosensor-assisted high-throughput screening method was designed and implemented to expedite the identification of high-performance PCA-producing strains. Finally, in a 5 L bioreactor, the final strain PCA05 achieved the highest PCA titer of 46.65 g/L, a yield of 0.23 g/g, and a productivity of 1.46 g/L/h for PCA synthesis from glucose using normal fed-batch fermentation. The strategies described herein serve as valuable guidelines for the production of other high-value and toxic products.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Metabolic engineering
Metabolic engineering 工程技术-生物工程与应用微生物
CiteScore
15.60
自引率
6.00%
发文量
140
审稿时长
44 days
期刊介绍: Metabolic Engineering (MBE) is a journal that focuses on publishing original research papers on the directed modulation of metabolic pathways for metabolite overproduction or the enhancement of cellular properties. It welcomes papers that describe the engineering of native pathways and the synthesis of heterologous pathways to convert microorganisms into microbial cell factories. The journal covers experimental, computational, and modeling approaches for understanding metabolic pathways and manipulating them through genetic, media, or environmental means. Effective exploration of metabolic pathways necessitates the use of molecular biology and biochemistry methods, as well as engineering techniques for modeling and data analysis. MBE serves as a platform for interdisciplinary research in fields such as biochemistry, molecular biology, applied microbiology, cellular physiology, cellular nutrition in health and disease, and biochemical engineering. The journal publishes various types of papers, including original research papers and review papers. It is indexed and abstracted in databases such as Scopus, Embase, EMBiology, Current Contents - Life Sciences and Clinical Medicine, Science Citation Index, PubMed/Medline, CAS and Biotechnology Citation Index.
期刊最新文献
Unraveling productivity-enhancing genes in Chinese hamster ovary cells via CRISPR activation screening using recombinase-mediated cassette exchange system. The faucet knob effect of DptE crotonylation on the initial flow of daptomycin biosynthesis. Versatile Xylose and Arabinose Genetic Switches development for Yeasts. Not all cytochrome b5s are created equal: How a specific CytB5 boosts forskolin biosynthesis in Saccharomyces cerevisiae Applying metabolic control strategies to engineered T cell cancer therapies
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1