Synthetic and Systems Biotechnology最新文献_第7页

Combination of enzyme engineering and quorum sensing system for efficient de novo biosynthesis of β-arbutin in Escherichia coli 结合酶工程和群体感应系统在大肠杆菌中高效从头合成β-熊果苷

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-08-22 DOI: 10.1016/j.synbio.2025.08.010

Zhuoyuan Liu , Dongming Liu , Lihao Deng , Kai LingHu , Xiaoyu Shan , Jingwen Zhou , Weizhu Zeng

{"title":"Combination of enzyme engineering and quorum sensing system for efficient de novo biosynthesis of β-arbutin in Escherichia coli","authors":"Zhuoyuan Liu , Dongming Liu , Lihao Deng , Kai LingHu , Xiaoyu Shan , Jingwen Zhou , Weizhu Zeng","doi":"10.1016/j.synbio.2025.08.010","DOIUrl":"10.1016/j.synbio.2025.08.010","url":null,"abstract":"<div><div>β-Arbutin is a plant-derived glycoside with various biological activities, which has been applied in pharmaceuticals and whitening cosmetics. Uneven carbon flux distribution and the low catalytic efficiency of arbutin synthase limit the biosynthesis of β-arbutin in microorganisms. In addition, commonly used high-cost, or toxic, inducible expression agents are not practical to use in industrial production. In this study, a biosynthetic pathway for β-arbutin was constructed in <em>Escherichia coli</em>. The competition and degradation pathways were knocked out, and the key pathway enzymes were identified and overexpressed, to increase β-arbutin production. Furthermore, by combining modifications to the phosphoketolase pathway and the acetic acid pathway, the carbon flux was redirected to the shikimic acid pathway. The substrate access channel of the arbutin synthase active site was modified by protein engineering, resulting in the R241H mutant, which increased β-arbutin production by 16.7 %, compared with wild type (WT). Finally, the luxR-based quorum sensing (QS) system was used to regulate the allocation of metabolic fluxes between cell growth and product synthesis, achieving a self-induced regulation of <em>E. coli,</em> without requiring exogenous inducers. The final engineered strain produced 81.9 g/L of β-arbutin from a fed-batch fermentation in a 5 L fermenter and the yield from glucose reached 0.29 g/g. This study presents a systematic strategy that can provide a reference for the construction of β-arbutin microbial cell factories.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 1-9"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144933820","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

Putative membrane protein Cgl2168 and its variant enhance thermotolerance in Corynebacterium glutamicum 推测膜蛋白Cgl2168及其变体增强谷氨酸棒状杆菌的耐热性

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-09-25 DOI: 10.1016/j.synbio.2025.09.018

Ning Gao , Xiaomeng Ni , Jiao Liu , Jiuzhou Chen , Yu Wang , Ping Zheng , Jibin Sun

{"title":"Putative membrane protein Cgl2168 and its variant enhance thermotolerance in Corynebacterium glutamicum","authors":"Ning Gao , Xiaomeng Ni , Jiao Liu , Jiuzhou Chen , Yu Wang , Ping Zheng , Jibin Sun","doi":"10.1016/j.synbio.2025.09.018","DOIUrl":"10.1016/j.synbio.2025.09.018","url":null,"abstract":"<div><div><em>Corynebacterium glutamicum</em> ATCC 13032 is widely used as the chassis strain for industrial production of various amino acids, but its thermotolerance limits productivity under high temperatures. Comparative genomic analysis in this study identified <em>cgl2168</em> as a key determinant of enhanced thermotolerance in a laboratory strain (13032-JN) compared to another stock (13032-TJ). A guanine at Residue 270 (G270) deletion in <em>cgl2168</em> (<em>cgl2168</em><sup>Δ270G</sup>) in 13032-JN significantly improved growth at 40 °C, while reverting this mutation or deleting <em>cgl2168</em> impaired thermotolerance. Structural analysis revealed the deletion extends the C-terminus with an additional α-helix. Transcriptomic profiling showed <em>cgl2168</em><sup>Δ270G</sup> increased gene expressions involved in oxidative phosphorylation (e.g., <em>atpABCDEFGH</em>), nitrate metabolism (<em>narIJHK</em>), some heat shock protein genes (e.g., <em>dnaK</em> and <em>groES</em>) and deletion of <em>cgl2168</em> resulted in upregulating sulfur metabolism (sulfonate transporter protein genes <em>ssuBCD</em>, sulfur-containing amino acid <em>cys</em> and <em>met</em> operons) and central carbon metabolism (<em>gltA</em>, <em>pyc</em>), while downregulating gluconeogenesis (<em>pck</em>) and fermentation (<em>ldh</em>) genes. These changes enhanced ATP synthesis, carbon utilization and oxidative tolerance. Predicted interactors of Cgl2168 linked it to respiratory chain function, stress responses, and cell envelope biogenesis. Thus, Cgl2168 and its variant Cgl2168<sup>Δ270G</sup> enhance thermotolerance by coordinating energy production, carbon metabolism, and stress adaptation, aiding industrial applications.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 287-297"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145325258","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

Competitive binding of TeXlnR and TeAmyR regulates cellulase production in Talaromyces endophyticus TeXlnR和TeAmyR的竞争结合调控了Talaromyces内生纤维素酶的产生

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-11-03 DOI: 10.1016/j.synbio.2025.10.010

Yixin Sun , Qi Sun , Tiantian Gao , Shumin Ren , Xiaomei Hu

{"title":"Competitive binding of TeXlnR and TeAmyR regulates cellulase production in Talaromyces endophyticus","authors":"Yixin Sun , Qi Sun , Tiantian Gao , Shumin Ren , Xiaomei Hu","doi":"10.1016/j.synbio.2025.10.010","DOIUrl":"10.1016/j.synbio.2025.10.010","url":null,"abstract":"<div><div>Efficient cellulase production is essential for its application in green bioprocessing, yet the transcriptional regulation of cellulase biosynthesis in <em>Talaromyces</em> sp. remains largely uncharacterized. In this study, we elucidate the regulatory functions of TeXlnR and TeAmyR in <em>Talaromyces endophyticus</em>. Our results demonstrate that TeXlnR, as a potent transcriptional activator, enhances enzyme production by over 33 %. RNA-seq analysis shows that <em>TeXlnR</em> overexpression upregulates 33 cellulase genes and 11 hemicellulase genes, while concurrently repressing the expression of the transcription factor <em>TeAmyR</em>. Further investigation identified TeAmyR as a repressor of cellulase production in <em>T. endophyticus</em>. EMSA and yeast one-hybrid analyses confirmed the specific competitive binding of these two transcription factors to the promoter regions of cellulase genes <em>eg5B</em> and <em>eg12A</em>. TeXlnR both directly activates target gene transcription and indirectly promotes cellulase synthesis by downregulating <em>TeAmyR</em> expression to reduce its competitive binding to promoters. These findings provide direct evidence for competitive regulation by transcription factors in <em>Talaromyces</em> sp. and establish a theoretical foundation for the rational engineering of industrial strains with improved cellulase production.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 356-369"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145465807","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

Harnessing prokaryotic amino acid transporters for metabolic engineering: mechanisms and biotechnological applications 利用原核氨基酸转运体进行代谢工程：机制和生物技术应用

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-10-22 DOI: 10.1016/j.synbio.2025.10.007

Junyao Huang , Wenming Wu , Zhicheng Fu , Chunyu Chao , Chongde Lai , Xiaoyu Wu , Hui Yang , Xiaohe Chu , Bangce Ye , Bin Zhang

{"title":"Harnessing prokaryotic amino acid transporters for metabolic engineering: mechanisms and biotechnological applications","authors":"Junyao Huang , Wenming Wu , Zhicheng Fu , Chunyu Chao , Chongde Lai , Xiaoyu Wu , Hui Yang , Xiaohe Chu , Bangce Ye , Bin Zhang","doi":"10.1016/j.synbio.2025.10.007","DOIUrl":"10.1016/j.synbio.2025.10.007","url":null,"abstract":"<div><div>As fundamental bioactive molecules and building blocks of cellular structures, amino acids play a crucial role in sustaining life, reflected in a global market valued at tens of billions of dollars. Recently, microbial fermentation has become a leading alternative to chemical synthesis and enzymatic catalysis for amino acid production, driving the need for robust strains. Efficient transport of amino acids out of the cell is essential for strain development, as it mitigates intracellular accumulation, cytotoxicity, and feedback inhibition, thereby improving fermentation efficiency. This review summarizes the physiological characteristics of amino acid transporters and their applications in the metabolic engineering of high-performance microbial strains. In particular, it provides a comprehensive overview of how amino acid transport systems are applied to develop amino acid-producing strains by enhancing substrate uptake, increasing precursor supply, directly promoting amino acid transportation, and reducing flux through competing metabolic pathways. Furthermore, this review explores future challenges and prospects in the field, offering theoretical guidance for engineering robust microbial platforms capable of producing not only amino acids but also high-value bioproducts.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 342-355"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145416571","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

Efficient synthesis of l-DOPA in Escherichia coli via cofactor and enzyme engineering 利用辅助因子和酶工程在大肠杆菌中高效合成左旋多巴

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-09-12 DOI: 10.1016/j.synbio.2025.09.011

Lihao Deng , Jurong Ping , Zhuoyuan Liu , Kai Linghu , Heng Zhang , Xiaoyu Shan , Weizhu Zeng , Jianghua Li , Jingwen Zhou

{"title":"Efficient synthesis of l-DOPA in Escherichia coli via cofactor and enzyme engineering","authors":"Lihao Deng , Jurong Ping , Zhuoyuan Liu , Kai Linghu , Heng Zhang , Xiaoyu Shan , Weizhu Zeng , Jianghua Li , Jingwen Zhou","doi":"10.1016/j.synbio.2025.09.011","DOIUrl":"10.1016/j.synbio.2025.09.011","url":null,"abstract":"<div><div>The global incidence of Parkinson's disease continues to rise. Levodopa (<span>l</span>-DOPA) is the core therapeutic drug, and efficient and sustainable production methods are needed. However, the complex metabolic pathways and the low catalytic efficiency of enzymes limit biosynthesis of <span>l</span>-DOPA in microorganisms. To address this issue, this study significantly enhanced the production efficiency of <span>l</span>-DOPA through a multi-dimensional, integrated metabolic and enzyme engineering approach. Firstly, the <em>de novo</em> synthesis pathway for <span>l</span>-DOPA was established through optimization of the promoter, ribosome-binding site (RBS), plasmid copy number, and tighly accurately regulating the expression level of key enzymes. Secondly, combined with metabonomic analysis, carbon metabolic flow was diverted, increasing the <span>l</span>-DOPA titer by 36.7 %. Glucose dehydrogenase (<em>BmgdH</em>) and gluconate kinase (<em>gntK</em>) were introduced to construct a cofactor regeneration system, which synergistically enhanced the supply of NADH and FADH<sub>2</sub>, increasing the <span>l</span>-DOPA conversion rate by 18 %. Next, the substrate tunnel of 4-hydroxyphenylacetic acid-3-monooxygenase subunit B (HpaB) was subjected to rational design, and mutant T292A significantly expanded the substrate channel, improved catalytic efficiency, and decreased <span>l</span>-tyrosine by 87 %. Finally, through the process optimization in a 5 L bioreactor (involving phased pH control and induction timing adjustment) achieved an <span>l</span>-DOPA titer of 60.73 g/L, the highest reported to date for <em>de novo</em> microbial synthesis. This research offers a novel approach for industrial biosynthesis of <span>l</span>-DOPA, and broadens engineering concepts for efficient synthesis of aromatic compounds.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 226-236"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145220363","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

Rapid detection of genotoxicity employing an engineered high-sensitivity and tight SOS-response biosensor in Escherichia coli MG1655 利用工程化的高灵敏度、紧密sos反应生物传感器快速检测大肠杆菌MG1655的遗传毒性

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-06-27 DOI: 10.1016/j.synbio.2025.06.011

Xin Yuan , Wenliang Hao , Chong Zhang , Yizhi Ji , Yi Wang

{"title":"Rapid detection of genotoxicity employing an engineered high-sensitivity and tight SOS-response biosensor in Escherichia coli MG1655","authors":"Xin Yuan , Wenliang Hao , Chong Zhang , Yizhi Ji , Yi Wang","doi":"10.1016/j.synbio.2025.06.011","DOIUrl":"10.1016/j.synbio.2025.06.011","url":null,"abstract":"<div><div>Industrial wastewater and pesticide residues contain numerous genotoxic compounds that pose serious health risks by causing DNA damage and potentially triggering carcinogenesis. Conventional analytical methods such as chromatography and mass spectrometry are limited by high costs, technical complexity, and lengthy procedures, making them unsuitable for rapid, large-scale screening. To address this need, we developed a fluorescence-based SOS-responsive whole-cell biosensor (SRWCB) by integrating the P<sub><em>sulA</em></sub> promoter with superfolder GFP (sfGFP). Through directed evolution of the <em>lexA</em> binding motif in P<sub><em>sulA</em></sub>, we obtained an optimized variant (P55) with significantly enhanced sensitivity—being 8.5 times more sensitive than the commonly used P<sub><em>umuDC</em></sub> system—while maintaining rapid response kinetics. Further engineering via antisense transcription improved biosensor tightness without compromising induction efficiency. The final construct (SRWCB-55-J23119) successfully detected genotoxicants in industrial wastewater, demonstrating its practical applicability. This study presents a sensitive, rapid, and robust biosensor platform for high-throughput environmental genotoxicity monitoring.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 117-126"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145096563","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

Systems metabolic engineering of dual pathways with stage-specific activation for high-level 5-aminolevulinic acid production in Escherichia coli 大肠杆菌生产高水平5-氨基乙酰丙酸的双途径与阶段特异性激活的系统代谢工程

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-09-08 DOI: 10.1016/j.synbio.2025.09.007

Yanghao Liu , Changgeng Li , Yu Miao , Zhichao Chen , Tang'en Shi , Ling Ma , Siyu Zhao , Zichen Yu , Lanxiao Li , Qingyang Xu

{"title":"Systems metabolic engineering of dual pathways with stage-specific activation for high-level 5-aminolevulinic acid production in Escherichia coli","authors":"Yanghao Liu , Changgeng Li , Yu Miao , Zhichao Chen , Tang'en Shi , Ling Ma , Siyu Zhao , Zichen Yu , Lanxiao Li , Qingyang Xu","doi":"10.1016/j.synbio.2025.09.007","DOIUrl":"10.1016/j.synbio.2025.09.007","url":null,"abstract":"<div><div>5-Aminolevulinic acid (5-ALA) is an important non-proteinogenic amino acid with wide applications in agriculture and medicine. To achieve high-level microbial production, this study established a dual-pathway reconstruction strategy in <em>Escherichia coli</em> by integrating the endogenous C5 pathway with an inducible exogenous C4 pathway. Multi-copy overexpression of <em>gltX</em>, <em>hemA</em>, and <em>hemL</em>, combined with enhanced glutamate supply and the introduction of a non-oxidative glycolysis pathway, effectively increased the C5 pathway flux and carbon efficiency, while product toxicity was significantly alleviated by strengthening efflux mechanisms and oxidative stress tolerance. A quorum sensing–based regulatory system was employed to dynamically regulate <em>hemB</em> expression, thereby balancing cell growth and 5-ALA biosynthesis, and a controlled glycine feeding strategy specifically activated the C4 pathway during the later fermentation stage, forming a stage-specific dynamic activation mechanism. Promoter regulation of <em>sucC/sucD</em> expression and enhanced endogenous PLP biosynthesis further stabilized the C4 flux. Fed-batch fermentation in a 5 L bioreactor resulted in a final 5-ALA titer of 37.34 g/L, demonstrating the industrial potential of this systems metabolic engineering strategy combining dual pathways and stage-specific activation control.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 141-151"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145096567","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

Integrated transcriptomic and metabolomic analyses uncover novel genetic targets for enhanced l-tryptophan production in Corynebacterium glutamicum 整合转录组学和代谢组学分析揭示了谷氨酸棒状杆菌增强l-色氨酸生产的新遗传靶点

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-09-29 DOI: 10.1016/j.synbio.2025.09.020

Yufei Dong , Rongsheng Gao , Nan Qin , Kunyu Liu , Youmeng Liu , Zhen Chen

{"title":"Integrated transcriptomic and metabolomic analyses uncover novel genetic targets for enhanced l-tryptophan production in Corynebacterium glutamicum","authors":"Yufei Dong , Rongsheng Gao , Nan Qin , Kunyu Liu , Youmeng Liu , Zhen Chen","doi":"10.1016/j.synbio.2025.09.020","DOIUrl":"10.1016/j.synbio.2025.09.020","url":null,"abstract":"<div><div><em>Corynebacterium glutamicum</em> is a promising microbial chassis for the industrial production of <span>l</span>-tryptophan, an essential amino acid with diverse applications and high market value. In our previous work, we constructed an <span>l</span>-tryptophan-overproducing <em>C. glutamicum</em> strain, TR26, through multiple rounds of rational metabolic engineering. Here, comparative transcriptomic and metabolomic analyses were conducted between TR26 and its progenitor MB001 to elucidate the underlying molecular mechanisms and identify potential bottlenecks for <span>l</span>-tryptophan production in TR26. Based on the differentially expressed genes identified, systematic gene overexpression and knockdown experiments led to the identification of two novel genetic targets, <em>glnK</em> and <em>sugR</em>. Specifically, the repression of <em>glnK</em> and overexpression of <em>sugR</em> in TR26 increased the <span>l</span>-tryptophan titer by 10.3 % and 16.5 % in fed-batch fermentation, and the yield by 7.2 % and 20.2 %, respectively. Further transcription profiling and intracellular metabolite analysis indicated that these improvements were associated with altered nitrogen metabolism, more efficient allocation of cellular resources, and enhanced supply of phosphoenolpyruvate (PEP), a key precursor in aromatic amino acid biosynthesis. This study expands our understanding of the regulation mechanisms governing <span>l</span>-tryptophan synthesis in <em>C. glutamicum</em> and provides valuable insights for further optimization of industrial cell factories.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 256-267"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145325256","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

Engineering mixed sugar metabolic channels in Pseudomonas putida to produce vanillic acid 改造恶臭假单胞菌混合糖代谢通道生产香草酸

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-10-09 DOI: 10.1016/j.synbio.2025.10.002

Peihan Wu , Haifeng Ding , Zhiqing Xu , Yan Zhang , Yanyan Dang , Bo Gao , Genlin Zhang

{"title":"Engineering mixed sugar metabolic channels in Pseudomonas putida to produce vanillic acid","authors":"Peihan Wu , Haifeng Ding , Zhiqing Xu , Yan Zhang , Yanyan Dang , Bo Gao , Genlin Zhang","doi":"10.1016/j.synbio.2025.10.002","DOIUrl":"10.1016/j.synbio.2025.10.002","url":null,"abstract":"<div><div>Hydrolysis of lignocellulosic biomass produces the mixed sugars of glucose (60–70 %), xylose (20–30 %), and arabinose (2–20 %), etc. Using mixed sugars instead of pure glucose for microbial biosynthesis will reduce the cost of carbon source and maximize utilization of biomass. However, carbon catabolite repression and poor adaptation of metabolic pathways are obstacles in the synergistic utilization of mixed sugars, thus resulting in low carbon utilization efficiency. Here, we engineered the mixed sugar metabolic channels in <em>Pseudomonas putida</em> to achieve their synergistic utilization for producing vanillic acid, a valuable aromatic compound with broad applications in the food, pharmaceuticals, cosmetics, and chemical industries. Expressing O-methyltransferase (<em>OMT</em>) and deleting vanillate-O-demethylase (<em>vanAB</em>) realized vanillic acid accumulation in <em>P. putida</em> from glucose. Introducing the xylose isomerase pathway enabled the strain to produce vanillic acid from xylose. Deleting glucose dehydrogenase (<em>gcd</em>) and transcriptional regulator (<em>hexR</em>), together expressing two critical pentose phosphate pathway enzymes (transketolase and transaldolase) effectively balanced glucose-xylose metabolic channels for vanillic acid production. Further assembling the arabinose oxidation pathway established the efficient metabolism of three sugars. The final engineered strain (VA12) produced 2.75 g/L vanillic acid in fed-batch fermentation with 20 g/L glucose, 10 g/L xylose and 10 g/L arabinose. This study effectively reduces carbon catabolite repression in the synergistic utilization of mixed sugars, and represents the first case utilizing glucose-xylose-arabinose for vanillic acid production, illustrating the capability of transforming lignocellulose hydrolyzate into valuable chemical products.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 277-286"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145325259","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

Identification of a novel β-1,4-galactosyltransferase from Haemophilus parainfluenzae for efficient biosynthesis of lacto-N-neotetraose in Escherichia coli 副流感嗜血杆菌β-1,4-半乳糖转移酶在大肠杆菌中高效合成乳酸-n -新四糖的鉴定

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-03-01 Epub Date: 2025-09-05 DOI: 10.1016/j.synbio.2025.09.006

Dan Liu , Chenhui Ying , Yingqi Ping , Shanquan Liang , Qiaojuan Yan , Zhengqiang Jiang

{"title":"Identification of a novel β-1,4-galactosyltransferase from Haemophilus parainfluenzae for efficient biosynthesis of lacto-N-neotetraose in Escherichia coli","authors":"Dan Liu , Chenhui Ying , Yingqi Ping , Shanquan Liang , Qiaojuan Yan , Zhengqiang Jiang","doi":"10.1016/j.synbio.2025.09.006","DOIUrl":"10.1016/j.synbio.2025.09.006","url":null,"abstract":"<div><div>As an important member of human milk oligosaccharides (HMOs), lacto-<em>N</em>-neotetraose (LNnT) has been recognized as a promising HMO with many physiological activities for infants. β-1,4-Galactosyltransferases play a vital role in the final step of LNnT biosynthesis in microbial cell factory. Herein, we identified a novel β-1,4-galactosyltransferase (Hpa-GalT) from <em>Haemophilus parainfluenzae</em> for the efficient biosynthesis of LNnT in <em>E. coli</em> BL21 star(DE3). After overexpression of the key genes in the synthetic pathway, the titer of LNnT reached 1.86 g/L in shake flask fermentation. Moreover, a variant A186P/E146Q of Hpa-GalT was obtained by molecular modification, resulting in an increase of the LNnT titer to 2.15 g/L. Subsequently, a series of genes related to competitive pathways were deleted, including <em>lacZ</em>, <em>ugd</em>, <em>wcaJ</em>, and <em>wecB</em>, the engineered strain produced a LNnT titer of 2.23 g/L. After optimization of LNnT transport along with LNT II utilization, the engineered strain AH31 produced 2.93 g/L of LNnT in shake flask fermentation. Furthermore, it produced a maximum titer of 32.6 g/L of total LNnT and 27.1 g/L of extracellular LNnT in a 5-L fermentor with the productivity for 0.52 g/L/h. Overall, this study provides a novel β-1,4-galactosyltransferase and lays a foundation for the efficient biosynthesis of LNnT.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 102-109"},"PeriodicalIF":4.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145061143","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}

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