ACS Publications最新文献_第6页

Synthesis and Characterization of a π-Extended Clar's Goblet. π-扩展型克拉氏杯的合成与表征。

IF 15.6 1区化学

Journal of the American Chemical Society Pub Date : 2025-10-20 DOI: 10.1021/jacs.5c07588

Shantanu Mishra, Manuel Vilas-Varela, Igor Rončević, Fabian Paschke, Florian Albrecht, Leo Gross, Diego Peña

{"title":"Synthesis and Characterization of a π-Extended Clar's Goblet.","authors":"Shantanu Mishra, Manuel Vilas-Varela, Igor Rončević, Fabian Paschke, Florian Albrecht, Leo Gross, Diego Peña","doi":"10.1021/jacs.5c07588","DOIUrl":"https://doi.org/10.1021/jacs.5c07588","url":null,"abstract":"Concealed non-Kekulé polybenzenoid hydrocarbons have no sublattice imbalance yet cannot be assigned a classical Kekulé structure, leading to an open-shell ground state with potential applications in organic spintronics. They constitute an exceedingly small fraction of the total number of polybenzenoid hydrocarbons that can be constructed for a given number of benzenoid rings, and their synthesis remains challenging. The archetype of such a system is the Clar's goblet (C38H18), a diradical proposed by Erich Clar in 1972 and recently synthesized on a Au(111) surface. Here, we report the synthesis of a π-extended Clar's goblet (C76H26), a tetraradical concealed non-Kekulé polybenzenoid hydrocarbon, by a combined in-solution and on-surface synthetic approach. By low-temperature scanning tunneling microscopy and atomic force microscopy, we characterized individual molecules adsorbed on a Cu(111) surface. We provide insights into the electronic properties of this elusive molecule, including the many-body nature of its ground and excited states, by mean-field and multiconfigurational quantum chemistry calculations.","PeriodicalId":49,"journal":{"name":"Journal of the American Chemical Society","volume":" ","pages":""},"PeriodicalIF":15.6,"publicationDate":"2025-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145327911","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Machine-Readable Structural Information Is Essential for Natural Products Research. 机器可读的结构信息对天然产物研究至关重要。

IF 3.6 2区生物学

Journal of Natural Products Pub Date : 2025-10-20 DOI: 10.1021/acs.jnatprod.5c00836

Karin Steffen, Nicholas H Oberlies, Antonis Rokas

引用次数: 0

Machine Learning Modeling for ABC Transporter Efflux and Inhibition: Data Curation, Model Development, and New Compound Interaction Predictions. ABC转运体外排和抑制的机器学习建模：数据管理，模型开发和新的化合物相互作用预测。

IF 4.5 2区医学

Molecular Pharmaceutics Pub Date : 2025-10-20 DOI: 10.1021/acs.molpharmaceut.5c01065

Nada J Daood, Sean R Carey, Elena Chung, Tong Wang, Anna Kreutz, Mounika Girireddy, Suman Chakravarti, Nicole C Kleinstreuer, Jacqueline B Tiley, Lauren M Aleksunes, Hao Zhu

{"title":"Machine Learning Modeling for ABC Transporter Efflux and Inhibition: Data Curation, Model Development, and New Compound Interaction Predictions.","authors":"Nada J Daood, Sean R Carey, Elena Chung, Tong Wang, Anna Kreutz, Mounika Girireddy, Suman Chakravarti, Nicole C Kleinstreuer, Jacqueline B Tiley, Lauren M Aleksunes, Hao Zhu","doi":"10.1021/acs.molpharmaceut.5c01065","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.5c01065","url":null,"abstract":"In recent years, multiple computational studies have used machine learning models to predict substrate binding and inhibition of ATP-binding cassette (ABC) transporters. However, many of these studies relied on relatively small training sets with limited applicability. In this study, we manually curated over 24,000 bioactivity records (i.e., inhibition, binding affinity, permeability) for the ABC transporters P-gp, BCRP, MRP1, and MRP2 from more than 900 literature sources in ChEMBL, with additional data from PubChem and Metrabase. This effort yielded eight data sets, comprising around 8800 unique chemicals with one or more substrate binding or inhibition activities for these four efflux transporters. Quantitative structure-activity relationship (QSAR) models were developed for each of the eight data sets using combinations of four machine learning algorithms and three sets of chemical descriptors. The resulting models demonstrated excellent performance by 5-fold cross-validation, achieving an average correct classification rate (CCR) of 0.764 for the substrate binding models and 0.839 for the inhibition models. Models were validated with additional compounds from DrugBank that were known substrates or inhibitors. We further analyzed how model predictions for efflux transporter activity could estimate exposure of the brain to xenobiotics. Notably, compounds predicted as P-gp and BCRP substrates were twice or more likely to have low brain exposure compared to compounds with high brain exposure. This study provides a large and curated drug transporter binding and inhibition database for computational modeling. Applicable models based on this large database for predicting transporter substrate binding and inhibition can be used to evaluate more complex drug bioactivities, such as exposure of protected tissues to chemicals.","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145335989","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of Pyrimido Pyridazine Analogs through Increased Whole Cell Target Engagement of the Dihydropteroate Synthase Pterin Binding Site in Gram-Negative Bacteria. 通过增加革兰氏阴性菌中二氢蝶呤合酶蝶呤结合位点的全细胞靶标接合来开发嘧啶吡啶嘧啶类似物。

IF 3.8 2区医学

ACS Infectious Diseases Pub Date : 2025-10-20 DOI: 10.1021/acsinfecdis.5c00635

Hannah E Snoke, Stephanie M Reeve, Suresh Dharuman, Miranda J Wallace, Victoria C Loudon, Ying Zhao, John J Bowling, Patricia A Murphy, Brett Waddell, Robin B Lee, Jürgen B Bulitta, Richard E Lee

{"title":"Development of Pyrimido Pyridazine Analogs through Increased Whole Cell Target Engagement of the Dihydropteroate Synthase Pterin Binding Site in Gram-Negative Bacteria.","authors":"Hannah E Snoke, Stephanie M Reeve, Suresh Dharuman, Miranda J Wallace, Victoria C Loudon, Ying Zhao, John J Bowling, Patricia A Murphy, Brett Waddell, Robin B Lee, Jürgen B Bulitta, Richard E Lee","doi":"10.1021/acsinfecdis.5c00635","DOIUrl":"https://doi.org/10.1021/acsinfecdis.5c00635","url":null,"abstract":"Dihydropteroate synthase (DHPS) is a critical enzyme in the folate biosynthetic pathway of bacteria, fungi, and protozoans. Sulfonamides successfully target the p-aminobenzoic acid (pABA) binding site of DHPS, forming a false product that obstructs the formation of 7,8-dihydropteroate and disrupts subsequent reactions in the pathway. Pyrimido[4,5-c]pyridazine-based inhibitors target the pterin binding site of DHPS, demonstrating high target affinity but minimal antimicrobial activity, which has previously been attributed to poor permeability without detailed analysis. In this study, we investigate the permeability limitations of our pyrimido pyridazine series in Gram-negative bacteria within the context of whole cell target engagement and cellular accumulation. To evaluate their whole cell target engagement against Escherichia coli DHPS (EcDHPS), we developed a robust luminescence-based HiBiT cellular thermal shift assay and combined it with surface plasmon resonance and an LC-MS/MS-based accumulation assay. This orthogonal assay platform was used to reevaluate the SAR of our Legacy pyrimido pyridazine compound series against EcDHPS and to facilitate the design of an exploratory series of compounds with improved permeability. From this series, we found that the removal or replacement of the negatively charged carboxylic acid pyrimido pyridazine side chain with a thiotetrazole or a nitrile group resulted in increased accumulation, improved whole cell target engagement, and moderate antimicrobial activity against E. coli.","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336033","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mechanistic Insights into How Juxtamembrane Residue Modulation Leads To LAMP2A Inactivation in Chaperone-Mediated Autophagy. 在伴侣介导的自噬中，近膜残基调节如何导致LAMP2A失活的机制见解。

IF 2.9 2区化学

The Journal of Physical Chemistry B Pub Date : 2025-10-19 DOI: 10.1021/acs.jpcb.5c06192

Ishwar Patel, Nidhi Malhotra

{"title":"Mechanistic Insights into How Juxtamembrane Residue Modulation Leads To LAMP2A Inactivation in Chaperone-Mediated Autophagy.","authors":"Ishwar Patel, Nidhi Malhotra","doi":"10.1021/acs.jpcb.5c06192","DOIUrl":"https://doi.org/10.1021/acs.jpcb.5c06192","url":null,"abstract":"The lysosome-associated membrane protein type 2A (LAMP2A) is a critical mediator of chaperone-mediated autophagy (CMA), a selective degradation pathway essential for cellular homeostasis. The trimeric assembly of LAMP2A serves as an intermediate in forming higher-order oligomers that are responsible for substrate translocation and degradation. However, the molecular determinants of the trimeric stability remain poorly understood. We performed 30 μs of all-atom molecular dynamics simulations in a realistic lysosomal membrane environment to investigate how different protonation states of juxtamembrane histidine residues influence the structural dynamics of wild-type (WT) LAMP2A and compare these with an experimentally characterized inactive mutant, in which four charged residues were replaced by alanines. Comparative analyses reveal that in the WT protein, this region is stabilized through charged lipid nanoclusters and contributes to maintaining proper tilt angles and membrane anchoring of the monomeric and trimeric assemblies, while the mutant buries these residues within the lipid bilayer due to hydrophobic mismatch, leading to altered tilting, reduced dynamicity, disrupted oligomeric stability, altered lipid distribution, and membrane properties. We identified key interacting residues stabilizing the WT oligomeric state and demonstrated how their loss compromises trimeric assembly in the mutant. Our results offer mechanistic insights into how the disruption of juxtamembrane residues impairs CMA activity, with potential implications for CMA regulation.","PeriodicalId":60,"journal":{"name":"The Journal of Physical Chemistry B","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145327861","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Integrating OLINK Proteomics and Single-Cell Analysis Reveals that DCBLD2 Potentiates VEGFA-Driven Angiogenesis in Retinal Detachment with Choroidal Detachment. 整合OLINK蛋白质组学和单细胞分析揭示DCBLD2增强vegf驱动的血管生成视网膜脱离伴脉络膜脱离。

IF 3.6 2区生物学

Journal of Proteome Research Pub Date : 2025-10-19 DOI: 10.1021/acs.jproteome.5c00380

Qiuhong Wang, Xuan Chen, Zhifeng Wu

{"title":"Integrating OLINK Proteomics and Single-Cell Analysis Reveals that DCBLD2 Potentiates VEGFA-Driven Angiogenesis in Retinal Detachment with Choroidal Detachment.","authors":"Qiuhong Wang, Xuan Chen, Zhifeng Wu","doi":"10.1021/acs.jproteome.5c00380","DOIUrl":"https://doi.org/10.1021/acs.jproteome.5c00380","url":null,"abstract":"Rhegmatogenous retinal detachment with choroidal detachment (RRDCD) is a severe ophthalmologic condition whose molecular drivers remain unclear. Here, we integrated Olink proteomics of vitreous fluid from RRDCD and RRD patients (n = 20 each) with single-cell protein activity inference to uncover key pathogenic mechanisms. Proteomic analysis identified 110 statistically significant differentially expressed proteins (DEPs) revealing a significant upregulation of pro-inflammatory pathways, including TNF-α/NF-κB signaling in RRDCD. This multimodal analysis pinpointed DCBLD2 and VEGFA as central cooperating regulators. Functional validation in choroidal endothelial cells confirmed that DCBLD2 and VEGFA act synergistically to enhanced cell proliferation, migration, and angiogenic tube formation. Mechanistically, we demonstrate that DCBLD2 potentiates VEGFA-driven effects by increasing VEGFR2 phosphorylation and activating its downstream AKT and ERK1/2 signaling cascades. Our study reveals a novel synergistic axis where DCBLD2 amplifies VEGFA/VEGFR2 signaling to drive the pathological angiogenesis and inflammation characteristic of RRDCD. This work not only deepens our understanding of RRDCD pathogenesis but also establishes DCBLD2 and VEGFA as promising cooperative biomarkers and therapeutic targets for future clinical intervention.","PeriodicalId":48,"journal":{"name":"Journal of Proteome Research","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145327863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Preparing a Dual-Species In Vitro Biofilm Model for Testing Antibiofilm Efficacy. 制备双菌种体外生物膜模型检测抗生物膜效果。

IF 4.5 2区医学

Molecular Pharmaceutics Pub Date : 2025-10-19 DOI: 10.1021/acs.molpharmaceut.5c00798

Kelli Randmäe, Kairi Lorenz, Marta Putrinš, Tanel Tenson, Karin Kogermann

{"title":"Preparing a Dual-Species In Vitro Biofilm Model for Testing Antibiofilm Efficacy.","authors":"Kelli Randmäe, Kairi Lorenz, Marta Putrinš, Tanel Tenson, Karin Kogermann","doi":"10.1021/acs.molpharmaceut.5c00798","DOIUrl":"https://doi.org/10.1021/acs.molpharmaceut.5c00798","url":null,"abstract":"All wounds are contaminated, and there is a risk of developing an infection. Furthermore, most wounds contain biofilm and are contaminated by two bacteria, termed dual-species, or more bacteria, termed polybacterial biofilms. New antibacterial and antibiofilm wound care products are constantly being developed to combat this problem. There is a need to develop more biorelevant and reproducible models to test the efficacy of these wound care products. We used an electrospun (ES) gelatin-glucose matrix (Gel-Gluc) as an artificial skin substrate for dual-species biofilm formation using wound pathogens Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, combining them in pairs. When analyzing the biofilms, selective agars were used to differentiate various bacteria from one another while counting. The developed method supported the growth of dual-species biofilm that contained both bacteria up to 108 CFU/Gel-Gluc after 24 h. Over 48 h, there was a decrease in the number of S. aureus in the biofilms. Confocal microscopy imaging allowed monitoring of the location of bacteria in the Gel-Gluc and proved that different species were located closely together. ES polycaprolactone (PCL) fibrous wound dressings containing chloramphenicol (CAM) or ciprofloxacin (CIP), or their pristine analogs, were used to test the model. Both ES fibrous wound dressings were effective in preventing dual-species biofilm formation. PCL-CIP fibrous dressing was also effective in treating biofilms. The efficacy of treatment of E. coli varied in different dual-species combinations of E. coli. The developed dual-species biofilm model on artificial skin (Gel-Gluc) supported the successful growth of different bacterial combinations and proved to be suitable for testing the efficacy of ES fibrous wound dressings in preventing and treating biofilms.","PeriodicalId":52,"journal":{"name":"Molecular Pharmaceutics","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145327846","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Chipless Millifluidics Device for Rapid Fabrication of Hepatic Spheroids with Decellularized Liver Matrix. 无芯片微流体装置用于快速制备脱细胞肝基质肝球体。

IF 5.5 2区医学

ACS Biomaterials Science & Engineering Pub Date : 2025-10-19 DOI: 10.1021/acsbiomaterials.5c01058

Sourita Ghosh, Amit Ghosh, Falguni Pati, Suhanya Duraiswamy

{"title":"Chipless Millifluidics Device for Rapid Fabrication of Hepatic Spheroids with Decellularized Liver Matrix.","authors":"Sourita Ghosh, Amit Ghosh, Falguni Pati, Suhanya Duraiswamy","doi":"10.1021/acsbiomaterials.5c01058","DOIUrl":"https://doi.org/10.1021/acsbiomaterials.5c01058","url":null,"abstract":"Spheroids - simple 3D cellular aggregates, provide a superior model for simulating the in vivo complexity and architecture often lacking in traditional 2D cell cultures. They have numerous applications, including drug toxicity testing, disease modeling, tumor research, and regenerative medicine. However, existing spheroid preparation methods face significant challenges, particularly in achieving consistent shapes and sizes. Our work focuses on developing an affordable, user-friendly, and robust device for automated spheroid generation. This device uniquely utilizes decellularized extracellular liver matrix to encapsulate HepG2 cells, producing uniform liver spheroids as precise droplets in virgin coconut oil. The novelty of our approach lies in its ability to quickly and efficiently produce spheroids in large quantities, allowing for reproducible spheroid production while addressing the limitations of standard methods. By enhancing control over spheroid morphology and facilitating large-scale generation, our device promises to significantly advance the field of in vitro modeling and improve the reliability of experimental outcomes in various biomedical applications.","PeriodicalId":8,"journal":{"name":"ACS Biomaterials Science & Engineering","volume":" ","pages":""},"PeriodicalIF":5.5,"publicationDate":"2025-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145317956","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Point-of-Care Solid-Phase PCR in a Vertical Microfluidic Chip Integrated with All-Dielectric Nanostructured Metasurface for Highly Sensitive, Multiplexed Pathogen Detection. 基于全介电纳米结构超表面的垂直微流控芯片的即时固相PCR技术用于高灵敏度、多路病原体检测。

IF 9.1 1区化学

ACS Sensors Pub Date : 2025-10-19 DOI: 10.1021/acssensors.5c02435

Islam Seder, Leonid Beliaev, Rodrigo Coronel Téllez, Christian Anthon, Dhouha Grissa, Tao Zheng, Jan Gorodkin, Sanshui Xiao, Yi Sun

{"title":"Point-of-Care Solid-Phase PCR in a Vertical Microfluidic Chip Integrated with All-Dielectric Nanostructured Metasurface for Highly Sensitive, Multiplexed Pathogen Detection.","authors":"Islam Seder, Leonid Beliaev, Rodrigo Coronel Téllez, Christian Anthon, Dhouha Grissa, Tao Zheng, Jan Gorodkin, Sanshui Xiao, Yi Sun","doi":"10.1021/acssensors.5c02435","DOIUrl":"https://doi.org/10.1021/acssensors.5c02435","url":null,"abstract":"Multiplexed solid-phase polymerase chain reaction (SP-PCR) has emerged as an indispensable modality for concurrent amplification of multiple genetic loci within a singular reaction vessel, facilitating efficient molecular diagnostics. Nevertheless, SP-PCR has seldom been integrated into point-of-care diagnostic devices due to several technical challenges, such as bubble formation during PCR, long reaction time, and low fluorescence signals generated from the PCR products on a solid surface. To circumvent these constraints, we engineered a microfluidic chip comprising SP-PCR and nanophotonic enhancement to enable highly sensitive, high-throughput, and cost-efficient molecular diagnostics. The chip's vertical orientation integrates preloaded reagent chambers for sequential lysis, washing, elution, and amplification, driven by a synchronized stepper motor and air vacuum, achieving robust nucleic acid purification and reverse transcription-PCR, and enabling bubble-free, gravity-assisted fluid dynamics during the PCR thermocycling. Thermal cycling is expedited through a dual-heater configuration alternating at subsecond intervals, obviating active cooling and shortening the reaction time. All-dielectric nanostructured metasurface was incorporated beneath the PCR chamber, allowing for the facile immobilization of DNA arrays to conduct SP-PCR. Taking advantage of guided-mode resonance supported by the metasurface and the SP-PCR approaches permits multiplexed detection and achieves a detection limit of 10 copies/reaction, highlighting the platform's potential for point-of-care diagnostics, personalized medicine, and high-throughput pathogen surveillance. Facile fabrication and automation emphasize scalability for mass production and deployment and collectively represent an advancement in point-of-care diagnostics.","PeriodicalId":24,"journal":{"name":"ACS Sensors","volume":" ","pages":""},"PeriodicalIF":9.1,"publicationDate":"2025-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145327925","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Dynamic Pathway Regulation Using a Quorum-Sensing Circuit to Improve the Production of L-Homoserine in Escherichia coli. 利用群体感应电路提高大肠杆菌l -高丝氨酸产量的动态途径调控。

IF 3.9 2区生物学

ACS Synthetic Biology Pub Date : 2025-10-19 DOI: 10.1021/acssynbio.5c00364

Kun Niu, Miao Zhang, Yi-Ming Kong, Yi-Fan Zhao, Le-Tian Tan, Fa-Man Lu, Zhi-Qiang Liu, Yu-Guo Zheng

{"title":"Dynamic Pathway Regulation Using a Quorum-Sensing Circuit to Improve the Production of L-Homoserine in Escherichia coli.","authors":"Kun Niu, Miao Zhang, Yi-Ming Kong, Yi-Fan Zhao, Le-Tian Tan, Fa-Man Lu, Zhi-Qiang Liu, Yu-Guo Zheng","doi":"10.1021/acssynbio.5c00364","DOIUrl":"https://doi.org/10.1021/acssynbio.5c00364","url":null,"abstract":"L-Homoserine has diverse applications in the fields of agrochemicals, pharmaceuticals, and animal feed; therefore, microbial fermentation using engineered cell factories has attracted widespread and intense attention. In this study, the nonauxotrophic strain for higher L-homoserine production was developed based on the previously constructed strain Escherichia coli HS. First, key genes involved in the biosynthesis pathways of essential amino acids were replenished with different strategies to address the growth deficiencies. Subsequently, carbon flux through the l-aspartate to L-homoserine was amplified by thrA overexpression. Furthermore, the supply of NADPH and ATP was optimized to synergistically enhance L-homoserine biosynthesis. Finally, the quorum sensing (QS) system esaI/esaR from Pantoea stewartii was introduced to dynamically regulate the carbon flux of l-threonine biosynthesis. And the results indicated that optimizing the regulatory efficiency of the QS system triggered autonomous downregulation of thrB during the high-cell-density phase, achieving a balanced metabolic flux competition between the l-threonine and L-homoserine biosynthesis pathways. The QS-regulated strain E. coli HS27/PA-P7QS produced 101.81 g/L L-homoserine with a yield of 0.41 g/g glucose after 96 h of fermentation in a 5-L bioreactor. This study demonstrates the feasibility of applying the QS system in E. coli for metabolic flux control, thereby providing novel insights into the rational design of amino acid biosynthesis pathways.","PeriodicalId":26,"journal":{"name":"ACS Synthetic Biology","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145327844","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0