Metabolic engineering最新文献_第6页

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 : 2026-01-01 Epub 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":"2026-01-01","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

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 : 2026-01-01 Epub 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":"2026-01-01","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

Genome-wide CRISPR screening identifies genes in recombinant human embryonic kidney 293 cells for increased ammonia resistance 全基因组CRISPR筛选鉴定重组人胚胎肾293细胞中增加氨抗性的基因

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2026-01-01 Epub Date: 2025-10-21 DOI: 10.1016/j.ymben.2025.10.008

Sang Yoon Lee , Hyun Seung Kim , Yeon Gu Kim , Seunghyeon Shin , Seokchan Kweon , Jae Jun Lee , Gyun Min Lee

{"title":"Genome-wide CRISPR screening identifies genes in recombinant human embryonic kidney 293 cells for increased ammonia resistance","authors":"Sang Yoon Lee , Hyun Seung Kim , Yeon Gu Kim , Seunghyeon Shin , Seokchan Kweon , Jae Jun Lee , Gyun Min Lee","doi":"10.1016/j.ymben.2025.10.008","DOIUrl":"10.1016/j.ymben.2025.10.008","url":null,"abstract":"<div><div>Ammonia, a byproduct of glutamine metabolism, inhibits cell growth and reduces product yield and quality in mammalian cell culture. To identify novel genes associated with ammonia resistance, a genome-wide CRISPR knockout screening was conducted in monoclonal antibody (mAb)-producing human embryonic kidney 293 (HEK-mAb) cells using a virus-free, recombinase-mediated cassette exchange-based gRNA interrogation method. The knockout cell library was subcultured for five consecutive passages under 20 mM NH<sub>4</sub>Cl, enriching cells with a sgRNA that conferred a proliferation advantage under high-ammonia conditions. Next-generation sequencing analysis of the enriched population identified three target genes -<em>WNT3</em>, <em>TSPAN1</em>, and <em>CYHR1</em>-among 19,114 genes. Knockout of these genes in HEK-mAb cells resulted in a 1.33- to 1.56-fold increase in maximum viable cell concentration and a 1.28- to 1.58-fold increase in maximum mAb concentration under 20 mM NH<sub>4</sub>Cl. Notably, <em>WNT3</em> knockout maintained <em>N</em>-glycan galactosylation proportions of mAb despite ammonia stress. These findings highlight the effectiveness of genome-wide CRISPR knockout screening in identifying novel gene targets for ammonia-resistant HEK293 cell, offering a promising strategy for improving mAb production.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 184-193"},"PeriodicalIF":6.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145351710","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

Emerging hosts for metabolic engineering 代谢工程的新宿主。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2026-01-01 Epub Date: 2025-10-21 DOI: 10.1016/j.ymben.2025.10.009

Hal S. Alper

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 : 2026-01-01 Epub 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":"2026-01-01","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

MDG1-mediated transcriptional reprogramming enhances cellulase production and alters thermal activity in recombinant Saccharomyces cerevisiae mdg1介导的转录重编程增强了重组酿酒酵母菌的纤维素酶生产并改变了热活性。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2026-01-01 Epub Date: 2025-09-09 DOI: 10.1016/j.ymben.2025.09.002

Chun Wan , Xue-Qing Wang , Hou-Ru Yue , Ming-Ming Zhang , Akihiko Kondo , Riaan den Haan , Tomohisa Hasunuma , Kai Li , Xin-Qing Zhao

{"title":"MDG1-mediated transcriptional reprogramming enhances cellulase production and alters thermal activity in recombinant Saccharomyces cerevisiae","authors":"Chun Wan , Xue-Qing Wang , Hou-Ru Yue , Ming-Ming Zhang , Akihiko Kondo , Riaan den Haan , Tomohisa Hasunuma , Kai Li , Xin-Qing Zhao","doi":"10.1016/j.ymben.2025.09.002","DOIUrl":"10.1016/j.ymben.2025.09.002","url":null,"abstract":"<div><div>The budding yeast <em>Saccharomyces cerevisiae</em> is one of the most widely used microbial cell factories for heterologous protein and enzyme production. However, improving production efficiency and tailoring enzyme properties remain a major challenge. Here we identified <em>MDG1</em>, a gene involved in the pheromone signaling pathway, as a previously unrecognized regulator that significantly enhances cellulase production in recombinant yeast. Overexpression of <em>MDG1</em> significantly increased the extracellular activities of β-glucosidase I (BGLI), cellobiohydrolase I (CBHI), and endo-glycosidase II (EGII). Intriguingly, <em>MDG1</em> overexpression also altered the thermal activity profile of BGLI, shifting its peak activity from 50 °C to 37 °C—an inversion relative to the parental strain. Integrated transcriptome analyses revealed that <em>MDG1</em> regulates the expression of genes involved in the cell cycle and protein folding. Targeted modulation of key cell cycle regulators (<em>CLN1, PCL1, SWI5</em>) further improved BGLI activity, confirming their functional involvement. Secretome analysis and functional assays identified the disulfide isomerase Pdi1p as a key contributor to the enhanced enzyme performance at 37 °C. Our study reveals a novel role of <em>MDG1</em> in coordinating gene networks to improve enzyme activities and reshape enzymatic properties.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 24-34"},"PeriodicalIF":6.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145035700","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 : 2026-01-01 Epub 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":"2026-01-01","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

Industrial-scale production of Lacto-N-tetraose in engineered Bacillus subtilis via modular pathway optimization 通过模块化途径优化在工程枯草芽孢杆菌中工业化生产乳酸- n -四糖

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2026-01-01 Epub Date: 2025-10-30 DOI: 10.1016/j.ymben.2025.10.012

Chunhua Wang , Shaoru Hu , Shenglong Wang , Jing Du , Ziyi Zhao , Ziyao Zheng , Huihui Qiu , Xiang Ma , Jun Li , Hao Liu , Mingfeng Cao , Weixia Gao

{"title":"Industrial-scale production of Lacto-N-tetraose in engineered Bacillus subtilis via modular pathway optimization","authors":"Chunhua Wang , Shaoru Hu , Shenglong Wang , Jing Du , Ziyi Zhao , Ziyao Zheng , Huihui Qiu , Xiang Ma , Jun Li , Hao Liu , Mingfeng Cao , Weixia Gao","doi":"10.1016/j.ymben.2025.10.012","DOIUrl":"10.1016/j.ymben.2025.10.012","url":null,"abstract":"<div><div>Lacto-N-tetraose (LNT), an important human milk oligosaccharide with prebiotic benefits, was successfully produced de novo in <em>Bacillus subtilis</em>, establishing this Generally Recognized as Safe (GRAS) organism as a suitable platform for infant nutritional ingredients. A detailed enzyme screening identified three key enzymes: β-1,3-galactosyltransferase from <em>Pseudogulbenkiania ferrooxidans</em>, β-1,3-N-acetylglucosaminyltransferase from <em>Neisseria polysaccharea</em>, and β-galactoside permease (LacY) from E<em>scherichia coli</em>. The co-expression of these enzymes in strain BPPY enabled the first complete biosynthesis of LNT in <em>B</em>. <em>subtilis</em>, achieving a yield of 1.42 g/L in shake-flask cultures. Advanced metabolic engineering strategies, such as disrupting competing pathways, enhancing UDP-GlcNAc/Gal precursor flow, and optimizing heterologous pathways, led to the development of strain BPPY31, which produced 7.83 g/L of LNT, a 5.5-fold increase. To efficiently regulate carbon flux, a cost-effective CRISPR interference (CRISPRi) system was created to downregulate essential competing genes (<em>zwf</em>, <em>pfkA</em>, <em>murAB</em>). The engineered strain BD7 yielded 12.51 g/L in flask cultures and an impressive 80.48 g/L in 5-L fed-batch bioreactors, with only 4.43 g/L of the byproduct lacto-N-triose II, achieving lactose and glucose conversion rates of 92.25 % and 24 %, respectively. This study reports the highest documented LNT titer to date using a GRAS-compliant biomanufacturing platform characterized by precise metabolic regulation, scalability, and significant potential for industrial production of human milk oligosaccharides.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 232-242"},"PeriodicalIF":6.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145383266","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

Evolution-assisted engineering of formate assimilation via the formyl phosphate route in Escherichia coli 在大肠杆菌中通过磷酸甲酰基途径进行甲酸同化的进化辅助工程。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2026-01-01 Epub Date: 2025-10-15 DOI: 10.1016/j.ymben.2025.10.004

Jenny Bakker , Maximilian Boinot , Karin Schann , Jörg Kahnt , Timo Glatter , Tobias J. Erb , Maren Nattermann , Sebastian Wenk

{"title":"Evolution-assisted engineering of formate assimilation via the formyl phosphate route in Escherichia coli","authors":"Jenny Bakker , Maximilian Boinot , Karin Schann , Jörg Kahnt , Timo Glatter , Tobias J. Erb , Maren Nattermann , Sebastian Wenk","doi":"10.1016/j.ymben.2025.10.004","DOIUrl":"10.1016/j.ymben.2025.10.004","url":null,"abstract":"<div><div>The transition towards a sustainable bioeconomy requires the use of alternative feedstocks, with CO<sub>2</sub>-derived formate emerging as a promising candidate for industrial biotechnology. Despite its beneficial characteristics as a feedstock, microbial assimilation of formate is limited by the inefficiency of naturally evolved formate-fixing pathways. To overcome this limitation, synthetic formate reduction cascades could enable formate assimilation via formaldehyde, a key intermediate of several existing one carbon assimilation pathways. Recently, the formyl phosphate route, combining ATP-dependent activation of formate to formyl phosphate, followed by its reduction to formaldehyde, was developed through enzyme engineering and characterized <em>in vitro</em>. In this work, we successfully established the formyl phosphate route <em>in vivo</em> by developing a selection strategy that couples formate reduction to growth in a threonine/methionine auxotrophic <em>Escherichia coli</em>. Through adaptive laboratory evolution, we achieved formate-dependent growth via this novel pathway. Evolved strains were capable of growing robustly with formate concentrations between 20 mM and 100 mM with glucose in the co-feed. Genomic and proteomic analyses together with activity assays uncovered that formate activation was limiting <em>in vivo</em>. This discovery guided the rational engineering of a strain capable of efficient formate assimilation through the formyl phosphate route. By demonstrating that novel enzyme activities can link formate reduction to cell growth, our study shows how synthetic metabolic routes can be functionally established inside the cell, paving the way for the engineering of more complex synthetic pathways.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 208-217"},"PeriodicalIF":6.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145311455","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

Harnessing the MEP pathway for heterologous (+)-nootkatone biosynthesis in a green microalga under fine-tuned light and carbon regimes 利用MEP途径在微调光和碳制度下在绿色微藻中进行异源(+)-诺卡酮生物合成

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2026-01-01 Epub Date: 2025-10-27 DOI: 10.1016/j.ymben.2025.10.010

Merve Saudhof , Jona Brückner , Timo Sürmene , Anke Rattenholl , Thomas Baier , Olaf Kruse

{"title":"Harnessing the MEP pathway for heterologous (+)-nootkatone biosynthesis in a green microalga under fine-tuned light and carbon regimes","authors":"Merve Saudhof , Jona Brückner , Timo Sürmene , Anke Rattenholl , Thomas Baier , Olaf Kruse","doi":"10.1016/j.ymben.2025.10.010","DOIUrl":"10.1016/j.ymben.2025.10.010","url":null,"abstract":"<div><div>The high-value sesquiterpenoid (+)-nootkatone has important applications in food, agriculture, and pharmaceutical industries. Extraction from plant material, however, is technically challenging and inefficient due to inherent low concentrations in native sources. Over the last decade, the eukaryotic green microalga <em>Chlamydomonas reinhardtii</em> has emerged as a powerful alternative for heterologous terpenoid production, due to a natively high carbon flux through its MEP pathway. This study describes strategic fusion protein designs of different valencene and farnesyl pyrophosphate (FPP) synthases, which allowed efficient (+)-valencene biosynthesis in the <em>C. reinhardtii</em> cytosol and also found the algal chloroplast to be highly suitable for heterologous production. Successful co-expression of cytochrome P450 monooxygenases resulted in a two-step oxidation towards (+)-nootkatone at a comparably high conversion rate of 76 % and was independent of recombinant reductase activity. In addition, the 1-deoxyxylulose-5-phosphate synthase (DXS) was found to be rate-limiting for increased sesquiterpenoid production.</div><div>Currently available photobioreactors suffer from limitations in light availability, which can hinder phototrophic growth, especially at higher cell densities. <em>C. reinhardtii</em> harbours the ability to use acetic acid as a carbon source, and fine-tuned cultivation regimes under photo-, mixo-, and heterotrophic conditions were tested to optimize heterologous sesquiterpene production. Customized scale-up cultivations in 2.5 L showed efficient volumetric production of 148 mg/L under phototrophic conditions and a maximal gravimetric production of 76 mg/g<sub>CDW</sub> under heterotrophic cultivation regimes, which displays a first industrially relevant (+)-nootkatone production concept in a green cell factory.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"93 ","pages":"Pages 260-270"},"PeriodicalIF":6.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145383253","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