{"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}
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.
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.