利用丙二酰辅酶a供给途径工程技术培育高产蒽醌大肠杆菌菌株。

IF 3.9 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Takatoshi Suematsu, Manami Takama, Itsuki Tomita, Shumpei Asamizu, Takahiro Bamba, Tomohisa Hasunuma
{"title":"利用丙二酰辅酶a供给途径工程技术培育高产蒽醌大肠杆菌菌株。","authors":"Takatoshi Suematsu, Manami Takama, Itsuki Tomita, Shumpei Asamizu, Takahiro Bamba, Tomohisa Hasunuma","doi":"10.1021/acssynbio.5c00354","DOIUrl":null,"url":null,"abstract":"<p><p>Anthraquinones are valuable compounds that are traditionally used as natural pigments and have diverse pharmacological activities, including antimicrobial and anticancer effects. In this study, we aimed to enhance the production of 1,3,5-trihydroxyanthraquinone (AQ256) using <i>Escherichia coli</i> (<i>E. coli</i>) as a host. AQ256 is biosynthesized from eight malonyl-CoA molecules via the type II polyketide synthase pathway. However, previous studies have reported very low production levels of AQ256 in <i>E. coli</i> (approximately 2.5 mg/L), mainly because of limited malonyl-CoA availability. To address this, we introduced a heterologous malonate assimilation pathway and reinforced the endogenous malonyl-CoA biosynthesis pathway. An <i>E. coli</i> strain harboring AQ256 biosynthetic genes from <i>Photorhabdus laumondii</i> TTO1 produced only 1.3 mg/L AQ256. Upon introducing the malonate assimilation pathway and cultivating in malonate-supplemented Luria-Bertani medium, production increased to 3.8 mg/L. Further enhancement of the endogenous malonyl-CoA supply through the coexpression of pantothenate kinase and acetyl-CoA carboxylase resulted in strain AQ-04, which produced 12.3 mg/L AQ256. Optimization of cultivation conditions enabled AQ-04 to achieve 23.9 mg/L AQ256, a 9.6-fold increase compared to previous studies. Our results demonstrate that the combination of introducing a malonate assimilation pathway and enhancing native malonyl-CoA supply is a highly effective strategy for increasing malonyl-CoA availability. This approach is promising for the biosynthesis of a wide range of malonyl-CoA-derived compounds.</p>","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":""},"PeriodicalIF":3.9000,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of a High Anthraquinone-Producing <i>Escherichia coli</i> Strain Using Malonyl-CoA Supply Pathway Engineering.\",\"authors\":\"Takatoshi Suematsu, Manami Takama, Itsuki Tomita, Shumpei Asamizu, Takahiro Bamba, Tomohisa Hasunuma\",\"doi\":\"10.1021/acssynbio.5c00354\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Anthraquinones are valuable compounds that are traditionally used as natural pigments and have diverse pharmacological activities, including antimicrobial and anticancer effects. In this study, we aimed to enhance the production of 1,3,5-trihydroxyanthraquinone (AQ256) using <i>Escherichia coli</i> (<i>E. coli</i>) as a host. AQ256 is biosynthesized from eight malonyl-CoA molecules via the type II polyketide synthase pathway. However, previous studies have reported very low production levels of AQ256 in <i>E. coli</i> (approximately 2.5 mg/L), mainly because of limited malonyl-CoA availability. To address this, we introduced a heterologous malonate assimilation pathway and reinforced the endogenous malonyl-CoA biosynthesis pathway. An <i>E. coli</i> strain harboring AQ256 biosynthetic genes from <i>Photorhabdus laumondii</i> TTO1 produced only 1.3 mg/L AQ256. Upon introducing the malonate assimilation pathway and cultivating in malonate-supplemented Luria-Bertani medium, production increased to 3.8 mg/L. Further enhancement of the endogenous malonyl-CoA supply through the coexpression of pantothenate kinase and acetyl-CoA carboxylase resulted in strain AQ-04, which produced 12.3 mg/L AQ256. Optimization of cultivation conditions enabled AQ-04 to achieve 23.9 mg/L AQ256, a 9.6-fold increase compared to previous studies. Our results demonstrate that the combination of introducing a malonate assimilation pathway and enhancing native malonyl-CoA supply is a highly effective strategy for increasing malonyl-CoA availability. This approach is promising for the biosynthesis of a wide range of malonyl-CoA-derived compounds.</p>\",\"PeriodicalId\":26,\"journal\":{\"name\":\"ACS Synthetic Biology\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":3.9000,\"publicationDate\":\"2025-09-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"ACS Synthetic Biology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1021/acssynbio.5c00354\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Synthetic Biology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1021/acssynbio.5c00354","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0

摘要

蒽醌是传统上用作天然色素的有价值的化合物,具有多种药理活性,包括抗菌和抗癌作用。本研究旨在以大肠杆菌为宿主,提高1,3,5-三羟基蒽醌(AQ256)的产量。AQ256是由8个丙二酰辅酶a分子通过II型聚酮合成酶途径生物合成的。然而,先前的研究报道了大肠杆菌中AQ256的产生水平非常低(约2.5 mg/L),主要是因为丙二酰辅酶a的可用性有限。为了解决这一问题,我们引入了异源丙二酸同化途径,并加强了内源性丙二酰辅酶a生物合成途径。携带laumondii光habdus TTO1生物合成基因的大肠杆菌菌株仅产生1.3 mg/L的AQ256。引入丙二酸同化途径并在添加丙二酸的Luria-Bertani培养基中培养后,产量增加到3.8 mg/L。通过泛酸激酶和乙酰辅酶a羧化酶的共同表达进一步增强内源丙二酰辅酶a的供应,使菌株AQ-04产生12.3 mg/L的AQ256。优化培养条件使AQ-04的AQ256含量达到23.9 mg/L,比以往研究提高9.6倍。我们的研究结果表明,引入丙二酸同化途径和增加天然丙二酸辅酶a供应的结合是提高丙二酸辅酶a可用性的有效策略。该方法有望用于广泛的丙二酰辅酶a衍生物的生物合成。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of a High Anthraquinone-Producing Escherichia coli Strain Using Malonyl-CoA Supply Pathway Engineering.

Anthraquinones are valuable compounds that are traditionally used as natural pigments and have diverse pharmacological activities, including antimicrobial and anticancer effects. In this study, we aimed to enhance the production of 1,3,5-trihydroxyanthraquinone (AQ256) using Escherichia coli (E. coli) as a host. AQ256 is biosynthesized from eight malonyl-CoA molecules via the type II polyketide synthase pathway. However, previous studies have reported very low production levels of AQ256 in E. coli (approximately 2.5 mg/L), mainly because of limited malonyl-CoA availability. To address this, we introduced a heterologous malonate assimilation pathway and reinforced the endogenous malonyl-CoA biosynthesis pathway. An E. coli strain harboring AQ256 biosynthetic genes from Photorhabdus laumondii TTO1 produced only 1.3 mg/L AQ256. Upon introducing the malonate assimilation pathway and cultivating in malonate-supplemented Luria-Bertani medium, production increased to 3.8 mg/L. Further enhancement of the endogenous malonyl-CoA supply through the coexpression of pantothenate kinase and acetyl-CoA carboxylase resulted in strain AQ-04, which produced 12.3 mg/L AQ256. Optimization of cultivation conditions enabled AQ-04 to achieve 23.9 mg/L AQ256, a 9.6-fold increase compared to previous studies. Our results demonstrate that the combination of introducing a malonate assimilation pathway and enhancing native malonyl-CoA supply is a highly effective strategy for increasing malonyl-CoA availability. This approach is promising for the biosynthesis of a wide range of malonyl-CoA-derived compounds.

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
CiteScore
8.00
自引率
10.60%
发文量
380
审稿时长
6-12 weeks
期刊介绍: The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.
×
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
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
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 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信