Metabolic engineering最新文献

Metabolic and enzyme rewiring enables high-production of vanillin in unconventional yeast. 代谢和酶重组使非常规酵母的香兰素高产成为可能。

IF 8.4 1区生物学

Metabolic engineering Pub Date : 2025-10-08 DOI: 10.1016/j.ymben.2025.10.002

Yan Guo,Liyang Zhou,Wanshu Lai,Zhilan Qian,Haishuang Yu,Menghao Cai

{"title":"Metabolic and enzyme rewiring enables high-production of vanillin in unconventional yeast.","authors":"Yan Guo,Liyang Zhou,Wanshu Lai,Zhilan Qian,Haishuang Yu,Menghao Cai","doi":"10.1016/j.ymben.2025.10.002","DOIUrl":"https://doi.org/10.1016/j.ymben.2025.10.002","url":null,"abstract":"Vanillin is an aromatic flavor compound widely used in the food, pharmaceutical, and cosmetic industries. Microbial biosynthesis offers a sustainable alternative to traditional plant extraction and chemical synthesis; however, the susceptibility of vanillin to redox reactions and the weak enzyme activity in cells severely limit the vanillin production capacity by microbial biosynthesis. This study presents the first successful attempt at de novo synthesis of vanillin in the unconventional yeast Komagataella phaffii. The initial titer was quite low (0.5 mg/L), but removal of 14 endogenous oxidoreductases to block vanillin conversion resulted in an 11.1-fold improvement in vanillin production. The combination of pathway rewiring and cofactor (nicotinamide adenine dinucleotide phosphate [NADPH] and S-adenosylmethionine) regeneration redirected the metabolic flux toward vanillin synthesis and achieved a further 19.9-fold improvement in vanillin production. Rational rewiring of the rate-limiting enzyme, caffeic acid O-methyltransferase (NtCOMT), generated a dominant mutant NtCOMTN312A/H315N from 70 variants, which promoted activity by 49.7% and prevented intermediate accumulation. These strategies eventually enabled the co-coupling of de novo biosynthesis and caffeic acid conversion, achieving the highest reported production of vanillin (1055.9 mg/L) by K. phaffii fermentation in a bioreactor. These findings highlight the potential of unconventional yeast as a chassis host for aromatic aldehyde synthesis and the construction of a versatile microbial platform for the production of carbonyl compounds.","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"40 1","pages":""},"PeriodicalIF":8.4,"publicationDate":"2025-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145261529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Metabolic engineering of Acinetobacter baylyi ADP1 for efficient utilization of pentose sugars and production of glutamic acid. 贝氏不动杆菌ADP1代谢工程对戊糖的高效利用和谷氨酸的生产。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-10-07 DOI: 10.1016/j.ymben.2025.10.001

Jin Luo, Elena Efimova, Ville Santala, Suvi Santala

{"title":"Metabolic engineering of Acinetobacter baylyi ADP1 for efficient utilization of pentose sugars and production of glutamic acid.","authors":"Jin Luo, Elena Efimova, Ville Santala, Suvi Santala","doi":"10.1016/j.ymben.2025.10.001","DOIUrl":"https://doi.org/10.1016/j.ymben.2025.10.001","url":null,"abstract":"<p><p>Efficient utilization of pentose sugars is critical for advancing sustainable biomanufacturing using lignocellulose. However, many host strains capable of consuming glucose and lignin-derived monomers are unable to utilize pentose sugars. Here, we engineered Acinetobacter baylyi ADP1 for the utilization of D-xylose and L-arabinose. We first modelled different pentose utilization pathways using flux balance analysis to choose the most optimal pathway. A marker-free approach combining transformation and selection facilitated the integration of the pentose catabolic gene clusters of the selected Weimberg pathway into the A. baylyi genome, generating strains capable of efficiently utilizing both D-xylose and L-arabinose as sole carbon sources without any additional engineering or adaptation. For D-xylose, the cells achieved the highest growth rate (μ=0.73 h<sup>-1</sup>) reported to date for non-native hosts engineered for pentose utilization. For L-arabinose, a growth rate of μ=0.40 h<sup>-1</sup> was achieved, which also surpassed the growth rate on a native substrate of A. baylyi, glucose (μ=0.37 h<sup>-1</sup>). Importantly, pentose utilization occurred simultaneously with glucose utilization. We then applied metabolic flux analysis using <sup>13</sup>C labeled xylose to reveal D-xylose metabolism in the engineered strain. To demonstrate the potential for bioproduction, L-glutamate was selected as a target compound. Deletion of sucAB and gabT, and the overexpression of gdhA enabled L-glutamate production. With the engineered strain, a carbon yield of 34% during co-utilization with succinate and 70% via whole-cell catalysis using resting cells was achieved. Notably, L-glutamate production directly from industrially relevant hemicellulose hydrolysate was demonstrated. This study establishes a robust platform for pentose utilization and bioproduction in A. baylyi ADP1 and highlights the potential for metabolic optimization.</p>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":" ","pages":""},"PeriodicalIF":6.8,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145258527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Engineering a biosensor based high-throughput screening platform for high-yield caffeic acid production in Escherichia coli 设计一个基于生物传感器的高通量筛选平台，用于大肠杆菌高产咖啡酸的生产。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-09-30 DOI: 10.1016/j.ymben.2025.09.010

Daoguang Tian , Zhen Qin , Weilin Liu , Qinggele Caiyin , Weiguo Li , Guang-Rong Zhao , Jianjun Qiao

{"title":"Engineering a biosensor based high-throughput screening platform for high-yield caffeic acid production in Escherichia coli","authors":"Daoguang Tian , Zhen Qin , Weilin Liu , Qinggele Caiyin , Weiguo Li , Guang-Rong Zhao , Jianjun Qiao","doi":"10.1016/j.ymben.2025.09.010","DOIUrl":"10.1016/j.ymben.2025.09.010","url":null,"abstract":"<div><div>Caffeic acid (CA) is a valuable phenolic compound with wide applications in pharmaceuticals, food additives, and materials. However, its microbial production faces several challenges, including low heterologous enzyme activity and product toxicity. Here, we report the development of an integrated biosensor-driven high-throughput screening (HTS) platform for the efficient production of CA in <em>Escherichia coli</em>. We first identified and characterized CarR, a novel phenolic acid-responsive transcription factor from <em>Acetobacterium woodii</em>, and engineered it into a <em>p</em>-coumaric acid (<em>p</em>-CA) biosensor. Systematic optimization of the <em>p</em>-CA biosensor, resulting in reduced background, extended dynamic range and increased sensitivity. By coupling this biosensor with fluorescence-activated cell sorting, we established an efficient HTS platform that enabled the rapid selection of an improved FjTAL<sup>G85S</sup> mutant with a 6.85-fold enhancement in catalytic activity and a robust <em>p</em>-CA-producing strain (M5) with enhanced tolerance to <em>p</em>-CA and CA. Subsequent bottom-up metabolic engineering in strain CA8 achieved a CA titer of 9.61 g L<sup>−1</sup> in a 5-L bioreactor, the highest reported titer to date. Our work not only overcomes key bottlenecks in CA biosynthesis (low tyrosine ammonia-lyase activity, CA and <em>p</em>-CA cytotoxicity) but also provides a powerful tool to accelerate the engineering of microbial cell factories for the production of <em>p</em>-CA and other derived chemicals.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 128-144"},"PeriodicalIF":6.8,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145203458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Edible fungus Fusarium venenatum: advances, challenges, and engineering strategies for future food production 食用菌镰刀菌：未来食品生产的进展、挑战和工程策略

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-09-25 DOI: 10.1016/j.ymben.2025.09.009

Sheng Tong, Qiyu Qiu, Jiaying Gao, Jiali Yu, Yaobo Xu, Zhihua Liao

{"title":"Edible fungus Fusarium venenatum: advances, challenges, and engineering strategies for future food production","authors":"Sheng Tong, Qiyu Qiu, Jiaying Gao, Jiali Yu, Yaobo Xu, Zhihua Liao","doi":"10.1016/j.ymben.2025.09.009","DOIUrl":"10.1016/j.ymben.2025.09.009","url":null,"abstract":"<div><div>By 2050, the global population is projected to reach 9.7 billion, necessitating a 70 % increase in traditional agricultural output to meet growing demands. However, critical constraints are emerging as arable land and water resources approach their sustainable utilization thresholds. In this context, ensuring safe, efficient, and sustainable food production has become a pivotal issue intertwined with national economy and people's livelihood. Microbial manufacturing based on microbial chassis and synthetic biology technology represents a transformative approach to future food production. Notably, the edible filamentous fungus <em>Fusarium venenatum</em> serves as an ideal chassis for next-generation future food biomanufacturing. However, there has been a lack of systematic reviews specifically focusing on the development of synthetic biology tools, chassis engineering, and chassis applications for this strain. This paper systematically summarizes the latest significant progress, from the perspectives mentioned above, in the use of <em>F. venenatum</em> for future food biomanufacturing. Furthermore, it discusses potential development directions and challenges, and proposes some available strategies, intending to provide ideas and guidance for the further development of <em>F. venenatum</em>-based future food production systems.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 115-127"},"PeriodicalIF":6.8,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145156005","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Reusable and modular combinatorial libraries for iterative metabolic engineering of Saccharomyces cerevisiae 用于酿酒酵母迭代代谢工程的可重用和模块化组合库。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-09-19 DOI: 10.1016/j.ymben.2025.09.006

Philip Tinggaard Thomsen, Peter Gockel, Christina Vasileiou, Ingrid Mohr, Marc Cernuda Pastor, Irina Borodina

{"title":"Reusable and modular combinatorial libraries for iterative metabolic engineering of Saccharomyces cerevisiae","authors":"Philip Tinggaard Thomsen, Peter Gockel, Christina Vasileiou, Ingrid Mohr, Marc Cernuda Pastor, Irina Borodina","doi":"10.1016/j.ymben.2025.09.006","DOIUrl":"10.1016/j.ymben.2025.09.006","url":null,"abstract":"<div><div>Efficiently rewiring microbial metabolism for molecule production lies at the core of industrial metabolic engineering. Combinatorial libraries are useful for directing metabolism towards molecule production; however, their construction is labor-intensive, and their use in iterative strain engineering campaigns is often restricted by site-specific genomic integration. Here we present an automation-friendly framework for generating reusable and modular integration-based combinatorial libraries that can be used repeatedly to build high-performing strains. We apply this approach to engineer the production of betacyanins, a commonly used red food colorant extracted from beetroots, in <em>Saccharomyces cerevisiae</em>. Iterative implementation of combinatorial libraries targeting the betacyanin biosynthesis pathway (design space: ∼25,000), precursors (design space: ∼43,000), and cofactors (design space: ∼26,000) consistently improved pigment production by 1.2–5.7-fold per cycle over seven rounds of engineering. Sequencing of high-performing library isolates from each round revealed unique insights into betacyanin and yeast metabolism, <em>e.g.</em> we found strong evidence implicating the <em>S. cerevisiae</em> cytochrome <em>b</em>5 in heterologous red beet pigment production. Altogether, this study demonstrates a framework for combinatorial library engineering well-suited for accelerating the development of high-performing cell factories for industrial fermentation processes.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 100-114"},"PeriodicalIF":6.8,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145103543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

A designed hybrid pathway for efficient synthesis of D-pantothenate in E. coli 大肠杆菌高效合成d -泛酸酯的杂交途径设计

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-09-19 DOI: 10.1016/j.ymben.2025.09.007

Chenkai Cao , Jilong Wang , Mengzhen Nie , Jing Zhao , Yuchen Wang , Kechun Zhang

引用次数: 0

Development of a thermophilic l-arabinose-inducible system in Acetivibrio thermocellus (Clostridium thermocellum) 热细胞活动弧菌（Clostridium thermocellum）嗜热l-阿拉伯糖诱导体系的建立。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-09-19 DOI: 10.1016/j.ymben.2025.09.008

Fenghua Liu , Chao Chen , Ya-Jun Liu , Edward A. Bayer , Itzhak Mizrahi , Yingang Feng

{"title":"Development of a thermophilic l-arabinose-inducible system in Acetivibrio thermocellus (Clostridium thermocellum)","authors":"Fenghua Liu , Chao Chen , Ya-Jun Liu , Edward A. Bayer , Itzhak Mizrahi , Yingang Feng","doi":"10.1016/j.ymben.2025.09.008","DOIUrl":"10.1016/j.ymben.2025.09.008","url":null,"abstract":"<div><div>Inducible genetic operation systems constitute essential tools in microbial synthetic biology and metabolic engineering. However, inducible systems in non-model microbes, particularly thermophiles, are rarely reported. <em>Acetivibrio thermocellus</em> (previously termed <em>Clostridium thermocellum</em>), a representative strain of thermophilic non-model microbes, currently serves as a promising chassis organism in biorefinery. Although various genetic tools are available for <em>A. thermocellus</em>, superior thermophilic inducible systems are in high demand. In this study, we developed a thermostable <span>l</span>-arabinose-inducible system (ThermoARAi) in <em>A. thermocellus</em> by utilizing the inducible promoter P<sub>abnE</sub> and repressor AraR from <em>Geobacillus stearothermophilus</em> T-6. Through systematic promoter engineering and optimization of induction conditions using a thermostable β-glucuronidase as reporter, the system exhibited dynamic range improvement from a 5.4-fold induction to a 175-fold induction with negligible leakage. Furthermore, the ThermoARAi system was appropriate for use in metabolic engineering, as validated by its applications in whole-cell saccharification of cellulosic substrates and degradation of amorphous polyethylene terephthalate films. The ThermoARAi system significantly expands the genetic toolkit for precise gene expression modulation, metabolic engineering, and biotechnological applications in <em>A. thermocellus</em>. Importantly, this approach may also serve as a foundation for developing genetic tools in other Clostridia that play key roles in diverse ecosystems, including the gut.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 89-99"},"PeriodicalIF":6.8,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145103541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Multi-layered transcriptional and post-translational fine-tuning of metabolic pathways for overproduction of L-valine l -缬氨酸过量产生的代谢途径的多层转录和翻译后微调。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-09-18 DOI: 10.1016/j.ymben.2025.09.005

Xutao Lang , Wenwen Yu , Xuewen Zhu , Xianhao Xu , Yanfeng Liu , Jianghua Li , Guocheng Du , Jian Chen , Xueqin Lv , Long Liu

{"title":"Multi-layered transcriptional and post-translational fine-tuning of metabolic pathways for overproduction of L-valine","authors":"Xutao Lang , Wenwen Yu , Xuewen Zhu , Xianhao Xu , Yanfeng Liu , Jianghua Li , Guocheng Du , Jian Chen , Xueqin Lv , Long Liu","doi":"10.1016/j.ymben.2025.09.005","DOIUrl":"10.1016/j.ymben.2025.09.005","url":null,"abstract":"<div><div>L-valine is an essential amino acid widely used in the food, pharmaceutical, and animal feed industries. Currently, engineering microbial cell factories to produce L-valine from low-cost feedstocks has emerged as a leading strategy. However, there is still a lack of an L-valine-producing strain that simultaneously exhibits high titer, high yield, and high productivity. The metabolic engineering strategies reported for L-valine biosynthesis primarily rely on conventional unidimensional, transcriptional-level modifications, which limit fine-tuning and do not provide comprehensive, multi-layered regulation. In this study, we constructed an <em>Escherichia coli</em> hyperproducer of L-valine by multiplexed transcriptional and post-translational fine-tuning of metabolic pathways. Initially, the transcriptional repression in L-valine synthetic pathway was eliminated by using promoter engineering strategy. Then, the “push-pull-inhibit” and transport engineering strategies were used to improve L-valine accumulation, achieving a flask fermentation titer of 21.6 g/L and a productivity of 0.45 g/L/h. Subsequently, we rationally designed membraneless organelles (MLOs) to enable spatial regulation of metabolic biosynthesis, which enhanced the targeted recruitment of dihydroxy-acid dehydratase and branched-chain amino acid aminotransferase. This spatial reorganization led to a 95.6 % increase in productivity, reaching 0.88 g/L/h. Finally, the best-performing strain produced 90.6 g/L L-valine in a 3-L bioreactor at 28 h, with a yield of 0.48 g/g glucose and a productivity of 3.24 g/L/h. To the best of our knowledge, this represents the highest L-valine productivity achieved to date. Our strategy provides a practical and effective approach for advancing microbial amino acid biosynthesis by multi-layered transcriptional and post-translational regulation of metabolic pathways.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 80-88"},"PeriodicalIF":6.8,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145093553","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Improved arginine production in Escherichia coli by harnessing the intracellular citrulline 利用胞内瓜氨酸提高大肠杆菌精氨酸产量。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-09-13 DOI: 10.1016/j.ymben.2025.09.003

Qi Sheng , Shengyang He , Guangjie Liang , Gang Meng , Chunguang Zhao , Aiying Wei , Lining Gou , Jia Liu , Xiaomin Li , Jing Wu , Liming Liu

{"title":"Improved arginine production in Escherichia coli by harnessing the intracellular citrulline","authors":"Qi Sheng , Shengyang He , Guangjie Liang , Gang Meng , Chunguang Zhao , Aiying Wei , Lining Gou , Jia Liu , Xiaomin Li , Jing Wu , Liming Liu","doi":"10.1016/j.ymben.2025.09.003","DOIUrl":"10.1016/j.ymben.2025.09.003","url":null,"abstract":"<div><div>L-arginine is a high-value amino acid with widely utilized in the food, feed, and pharmaceutical industries. However, its large-scale biosynthesis remains limited by the low efficiency of current microbial strains. In this study, intracellular citrulline accumulation in <em>Escherichia coli</em>-Arg4 was enhanced by 2.45-, 1.90-, and 1.94-fold through supplementation with monosodium glutamate, monosodium aspartate, and glutamine hydrochloride, respectively. Correspondingly, L-arginine titers increased by 47.85 %, 21.18 %, and 10.66 %. Metabolic flux analysis and transcriptomic profiling indicated that exogenous ammonia donors redirected flux through critical metabolic nodes, including oxaloacetate, α-ketoglutarate, and citrulline, thus increasing precursor availability and enhancing L-arginine biosynthesis. Based on these findings, eight key gene targets, such as <em>gdhA</em>, <em>ppc</em>, <em>icd</em>, <em>aspC</em>, <em>glnA</em>, <em>pyrF</em>, <em>gltA</em>, and <em>argF</em> were identified for pathway optimization. Promoter engineering was subsequently employed to modulate their expression, and heterologous <em>gdhA</em> from <em>Salmonella enterica</em> and <em>glnA</em> from <em>Bacillus subtilis</em> were introduced. Consequently, an optimized strain, <em>E. coli</em>-Arg10, was constructed. Following process optimization in a 1000-L fermenter, the titer, yield and productivity of <em>E. coli</em>-Arg10 was achieved 108.33 g/L, 0.54 g/g, and of 2.26 g/L/h, respectively. These results highlight a scalable and efficient approach for microbial L-arginine production.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 46-59"},"PeriodicalIF":6.8,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145068229","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Systematic rewiring of Bacillus subtilis for efficient de novo biosynthesis of the neuroprotectant cytidine-5′-diphosphocholine 枯草芽孢杆菌系统重新布线，有效地重新合成神经保护剂胞苷-5'-二磷酸胆碱。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-09-11 DOI: 10.1016/j.ymben.2025.09.004

Shaomei Yang , Xu Feng , Hao Wei , Yanshuang Wang , Shouying Fu , Xiuzhen Gao , Qinyuan Ma

{"title":"Systematic rewiring of Bacillus subtilis for efficient de novo biosynthesis of the neuroprotectant cytidine-5′-diphosphocholine","authors":"Shaomei Yang , Xu Feng , Hao Wei , Yanshuang Wang , Shouying Fu , Xiuzhen Gao , Qinyuan Ma","doi":"10.1016/j.ymben.2025.09.004","DOIUrl":"10.1016/j.ymben.2025.09.004","url":null,"abstract":"<div><div>Cytidine-5′-diphosphocholine (CDP-choline) is a crucial neuroprotective agent. Current industrial production relies on chemical and enzymatic methods that face inherent sustainability challenges and share a dependence on the costly precursor, cytidine monophosphate (CMP). Here, we report the systems metabolic engineering of <em>Bacillus subtilis</em> for the efficient, <em>de novo</em> biosynthesis of CDP-choline from glucose, completely obviating the need for CMP. A synthetic pathway was first established by introducing heterologous choline kinase and phosphocholine cytidylyltransferase. Subsequently, a multi-module engineering strategy was implemented, focusing on enhancing precursor supply and redirecting carbon metabolism. This involved systematically optimizing choline uptake by overexpressing the transporter OpuD and deleting the transcriptional repressor <em>opcR</em>, fortifying the cytidine triphosphate (CTP) pool by overexpressing feedback-resistant CTP synthase gene <em>pyrG</em><sup>E156K</sup>, and deleting the transcriptional repressor <em>pyrR</em> along with other pyrimidine nucleotide consumption genes, and channeling carbon flux towards the TCA cycle by reducing pyruvate and malate consumption. The final engineered strain achieved a titer of 4.79 ± 0.24 g/L CDP-choline in a 5 L fed-batch bioreactor, with a high specific yield of 149.0 ± 5.8 mg/g DCW. Notably, the process exhibited a highly advantageous intracellular accumulation of 92.7 %, which simplifies downstream purification. This study represents the first successful demonstration of CDP-choline production from simple sugars in a microbial host, establishing a robust and economically competitive platform for its industrial manufacture.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 35-45"},"PeriodicalIF":6.8,"publicationDate":"2025-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145056657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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