Synthetic and Systems Biotechnology最新文献_第4页

Design principles for optogenetic-based targeted protein degradation 基于光遗传学的靶向蛋白降解设计原则

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2025-12-31 DOI: 10.1016/j.synbio.2025.12.006

Yunyue Chen , Siyifei Wang , Leiying Xie , Luhao Zhang , Min Zhu , Yingke Xu

{"title":"Design principles for optogenetic-based targeted protein degradation","authors":"Yunyue Chen , Siyifei Wang , Leiying Xie , Luhao Zhang , Min Zhu , Yingke Xu","doi":"10.1016/j.synbio.2025.12.006","DOIUrl":"10.1016/j.synbio.2025.12.006","url":null,"abstract":"<div><div>Precise regulation of protein abundance is essential for understanding dynamic cellular processes and for advancing therapeutic development. However, existing approaches lack the spatiotemporal resolution required to these cellular processes. Recent advances in optogenetics have enabled the design of optogenetic targeted protein degradation systems (Opto-TPD) allowing reversible and non-invasive control of protein stability with high spatiotemporal precision. In this review, we systematically summarize the design principles of Opto-TPD tools, including those based on light-oxygen-voltage (LOV)-domain conformational systems, light-inducible dimerization systems, and light-controlled degradation tool expression systems. We further highlight their applications in probing protein function, modulating signaling pathways, and therapeutic translations. By comparing the mechanistic features, performance, and limitations of each platform, we aim to provide a comprehensive resource for guiding future tool optimization. Altogether, these Opto-TPD tools represent a powerful and versatile complement to existing protein manipulation technologies, expanding the toolbox for precise control of protein homeostasis in living systems.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 255-264"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883629","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

Multi-gene Co-expression systems in E. coli: From single-vector designs to programmable expression platforms 大肠杆菌的多基因共表达系统：从单载体设计到可编程表达平台

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2026-01-10 DOI: 10.1016/j.synbio.2025.12.008

Rui Liu , Lu-Wei Wang , Zi-Han Gao , Xiao-Tong Sun , Shu-Ran Lv , Huan Liu , Sa-ouk Kang , Bo Sun

{"title":"Multi-gene Co-expression systems in E. coli: From single-vector designs to programmable expression platforms","authors":"Rui Liu , Lu-Wei Wang , Zi-Han Gao , Xiao-Tong Sun , Shu-Ran Lv , Huan Liu , Sa-ouk Kang , Bo Sun","doi":"10.1016/j.synbio.2025.12.008","DOIUrl":"10.1016/j.synbio.2025.12.008","url":null,"abstract":"<div><div><em>Escherichia coli</em> (<em>E. coli</em>) has long served as a versatile workhorse for recombinant protein production. As synthetic biology expands the demand for coordinated expression of multiple genes, co-expression systems in <em>E. coli</em> have evolved from basic dual-gene constructs to programmable, polygenic expression platforms. This review critically examines the major strategies enabling multigene co-expression in <em>E. coli,</em> including internal ribosome entry sites (IRES), 2A self-cleaving peptides, dual-promoter cassettes, multicistronic operons, and multi-plasmid configurations. We highlight the mechanistic principles, design trade-offs, and regulatory bottlenecks associated with each approach, such as translational imbalance, inclusion body formation, and plasmid compatibility. Real-world applications in metabolic engineering, complex protein assembly, and biomanufacturing are analyzed to demonstrate the functional advantages of these systems. Finally, we explore emerging programmable toolkits that integrate modular architecture, expression modeling, and AI-assisted design, paving the way for next-generation synthetic expression control in microbial chassis. This review offers a comprehensive and strategic roadmap for researchers engineering multi-gene systems in <em>E. coli</em> and beyond.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 330-341"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145938533","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

Precise l-threonine-to-l-isoleucine pathway regulation for engineering high-efficiency whole-cell biocatalysts 精确的l-苏氨酸到l-异亮氨酸途径调控工程高效全细胞生物催化剂

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2025-10-24 DOI: 10.1016/j.synbio.2025.10.009

Heng Zhang , Fuqiang Song , Ke Wang , Faqing Wu , Lihao Deng , Kun Qiu , Jingwen Zhou

{"title":"Precise l-threonine-to-l-isoleucine pathway regulation for engineering high-efficiency whole-cell biocatalysts","authors":"Heng Zhang , Fuqiang Song , Ke Wang , Faqing Wu , Lihao Deng , Kun Qiu , Jingwen Zhou","doi":"10.1016/j.synbio.2025.10.009","DOIUrl":"10.1016/j.synbio.2025.10.009","url":null,"abstract":"<div><div><span>l</span>-Isoleucine (L-Ile), a critical branched-chain amino acid with diverse applications in food, pharmaceutical, and cosmetic industries, is difficult to produce efficiently at scale in microbial systems due to metabolic bottlenecks and cofactor limitations. This study metabolically engineered <em>Escherichia coli</em> BL21(DE3) to develop a whole-cell biocatalyst for efficient L-Ile biosynthesis. Key strategies included screening acetohydroxy acid synthase (AHAS) isoenzymes, identifying <em>ilvGM</em>-encoded AHAS II as the optimal enzyme, relieving feedback inhibition of <em>ilvA</em> (encoding <span>l</span>-threonine dehydratase) through mutant screening, and optimizing genetic circuits (promoter tuning, plasmid copy number). Dual-precursor supplementation revealed <span>l</span>-threonine as a critical factor for suppressing <span>l</span>-valine byproduct. Fed-batch fermentation in a 5 L bioreactor achieved a peak molar conversion rate of 98.4 %, yielding 40.1 g/L L-Ile within 36 h. The mass conversion rate (L-Ile/glucose) achieved 0.36 g/g and the production efficiency achieved 1.11 g/L/h, demonstrating the feasibility of whole-cell catalysis. This work provides a robust framework for industrial L-Ile production and transferable strategies for branched-chain amino acid pathway optimization.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 1-9"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145449296","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

Truncation of LPD1 promoter and adaptive evolution increase cytosolic acetyl-CoA supply in yeast LPD1启动子的截断和适应性进化增加了酵母胞质乙酰辅酶a的供应

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2025-11-08 DOI: 10.1016/j.synbio.2025.10.013

Ling Qin , Shoujie He , Dan Yuan, Yuyang Pan, Zhibo Yan, Mingtao Huang

{"title":"Truncation of LPD1 promoter and adaptive evolution increase cytosolic acetyl-CoA supply in yeast","authors":"Ling Qin , Shoujie He , Dan Yuan, Yuyang Pan, Zhibo Yan, Mingtao Huang","doi":"10.1016/j.synbio.2025.10.013","DOIUrl":"10.1016/j.synbio.2025.10.013","url":null,"abstract":"<div><div>Acetyl-CoA is a central metabolic intermediate that serves as a key precursor for the biosynthesis of high-value compounds such as terpenoids. However, its compartmentalization within <em>Saccharomyces cerevisiae</em> limits its availability in the cytosol, constraining production of cytosol-derived metabolites. In this study, we aimed to redirect carbon flux toward cytosolic acetyl-CoA synthesis by reducing entry into the tricarboxylic acid cycle. To achieve this, we attenuated <em>LPD1</em> expression by deleting the noncoding RNA SUT526, which is located within the <em>LPD1</em> promoter region and overlaps an upstream regulatory element. This intervention impaired cell growth and hindered the utilization of non-fermentable carbon sources such as ethanol. To address this limitation, adaptive laboratory evolution was performed in ethanol-based medium, leading to rapid recovery of growth and extended cell viability. The evolved strains exhibited enhanced acetyl-CoA synthetase activity and elevated squalene production, suggesting an increased cytosolic acetyl-CoA supply. These improvements reflect enhanced flux through acetyl-CoA-dependent biosynthetic pathways. This work presents a targeted strategy for modulating central carbon metabolism to increase cytosolic acetyl-CoA supply, providing a framework for efficient production of acetyl-CoA derived compounds in yeast.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 10-19"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145475940","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

High-level production of vitamin K2 in Escherichia coli via modular molecular engineering 通过模块化分子工程在大肠杆菌中高水平生产维生素K2

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2025-11-12 DOI: 10.1016/j.synbio.2025.11.001

Zelin Lu , Zhongshi Huang , Zhengyin Wu , Zhengwen Zhu , Yibo Zhu , Xiaonuo Teng , Huyang Chen , Jingwen Zhou , Fuqiang Ma , Xinglong Wang

{"title":"High-level production of vitamin K2 in Escherichia coli via modular molecular engineering","authors":"Zelin Lu , Zhongshi Huang , Zhengyin Wu , Zhengwen Zhu , Yibo Zhu , Xiaonuo Teng , Huyang Chen , Jingwen Zhou , Fuqiang Ma , Xinglong Wang","doi":"10.1016/j.synbio.2025.11.001","DOIUrl":"10.1016/j.synbio.2025.11.001","url":null,"abstract":"<div><div>Menaquinone-7 (MK-7), a key form of vitamin K2 with wide-ranging nutritional and pharmaceutical applications, has attracted increasing interest for microbial production. Here, we developed an integrated modular metabolic engineering strategy in <em>Escherichia coli</em> to enhance MK-7 biosynthesis. Cellular membrane capacity and acetate metabolism were rewired to improve precursor supply for the mevalonate (MVA) pathway, while arabinose induction was applied to overexpress three critical enzymes, including BsHepPPS (<em>Bacillus subtilis</em>), EcMenA (<em>E. coli</em>), and BsUbiE (<em>B. subtilis</em>). Among them, EcMenA was identified as a major bottleneck. Rational protein engineering based on folding free energy analysis and consensus design yielded the EcMenA mutant G110W, which produced 102.55 mg/L MK-7 in shake-flask fermentation, a 57.2 % increase compared with the wild-type (WT) enzyme. Further active-site hotspot random mutagenesis generated a G110W-Q57T double mutant, raising MK-7 production to 176.38 mg/L, a 72 % increase compared to the single mutant. Optimization of EcMenA expression cassette by ribosome binding site redesign using a generative network further improved MK-7 titer to 227.53 mg/L in shake flasks. Finally, scale-up fermentation in a 50-L bioreactor, combined with optimized fermentation strategies, achieved a maximum MK-7 titer of 2.18 g/L. This study establishes a systematic framework integrating metabolic rewiring, enzyme engineering, and expression optimization, providing a robust platform for industrial-scale MK-7 production in microbial hosts.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 42-51"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145527394","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

Combining multiple metabolic strategies for efficient production of longifolene in Saccharomyces cerevisiae 结合多种代谢策略高效生产酿酒酵母长叶烯

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2025-11-18 DOI: 10.1016/j.synbio.2025.10.014

Yachao Xin , Jingping Du , Weiqiang Zhang , Haoran Bi , Limin Ba , Kai Wang , Yanhui Liu

{"title":"Combining multiple metabolic strategies for efficient production of longifolene in Saccharomyces cerevisiae","authors":"Yachao Xin , Jingping Du , Weiqiang Zhang , Haoran Bi , Limin Ba , Kai Wang , Yanhui Liu","doi":"10.1016/j.synbio.2025.10.014","DOIUrl":"10.1016/j.synbio.2025.10.014","url":null,"abstract":"<div><div>Longifolene is a sesquiterpene commonly found in the heavy turpentine oil of pine plants, with various applications ranging from pest control and fragrance production to synthetic biofuels. While <em>S. cerevisiae</em> cell factories can effectively accumulate longifolene, further optimization and refinement of the metabolic modifications are still needed to improve the yield and conversion efficiency of longifolene production. In this study, we first explored enzyme fusion technology to enhance the efficiency of longifolene synthase catalysis, and improved acetyl-CoA availability by adjusting the pyruvate bypass pathway and introducing the synthetic chimeric citrate lyase pathway. The introduction of the formate dehydrogenase module was also used to supplement reducing power. By combining these strategies, the yield of longifolene reached 78.637 mg/L in shake flasks and 2063.7 mg/L in a 5 L bioreactor through fed-batch cultivation. This is the highest reported yield of longifolene to date. This study has important fundamental significance for the construction of biosynthetic factories for longifolene and other terpenes.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 82-90"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145577446","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

Synthesis of tunable copolymers of 3-hydroxybutyrate and 3-hydroxyvalerate by engineered Halomonas bluephagenesis and their characterizations 工程蓝生盐单胞菌合成3-羟基丁酸酯和3-羟基戊酸酯可调共聚物及其表征

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2025-11-27 DOI: 10.1016/j.synbio.2025.11.007

Shaowei Li , Jinghui Wang , Yaoyao Zhang , Kaixin Du , Jiangnan Chen , Jianping Sun , Huan Wang , Pengfei Ouyang , Xuanming Xu , Fuqing Wu , Fang Yang , Guo-Qiang Chen

{"title":"Synthesis of tunable copolymers of 3-hydroxybutyrate and 3-hydroxyvalerate by engineered Halomonas bluephagenesis and their characterizations","authors":"Shaowei Li , Jinghui Wang , Yaoyao Zhang , Kaixin Du , Jiangnan Chen , Jianping Sun , Huan Wang , Pengfei Ouyang , Xuanming Xu , Fuqing Wu , Fang Yang , Guo-Qiang Chen","doi":"10.1016/j.synbio.2025.11.007","DOIUrl":"10.1016/j.synbio.2025.11.007","url":null,"abstract":"<div><div>Poly(3-hydroxybutyrate-<em>co</em>-3-hydroxyvalerate) (PHBV) with a 0–30 mol% controllable range of 3HV ratios was produced by <em>Halomonas bluephagenesis</em> (<em>H. bluephagenesis</em>) and characterized. An endogenous plasmid containing <em>scpA</em> and <em>scpB</em> encoding methylmalonyl-CoA epimerase and methylmalonyl-CoA decarboxylase, respectively, redirects succinyl-CoA toward propionyl-CoA, enabling <em>de novo</em> PHBV synthesis with a 1.7 mol% 3HV. Deletion of <em>sdhE</em> and <em>prpC</em> encoding succinate dehydrogenase and 2-methylcitrate synthase, respectively, further enhanced the 3HV to 4 mol%. <em>H. bluephagenesis</em> GZ05 was engineered for late-phase-specific MreB (a cytoskeletal protein) degradation, enlarged intracellular PHBV granules for enhanced PHBV synthesis, and convenient downstream. A series of growth experiments was conducted in a 7 L bioreactor fed with valerate to produce PHBV with various 3HV molar ratios (2–27 mol%). A quantitative relationship between valerate concentration and the final 3HV molar ratio was established with an R<sup>2</sup> = 0.9833, enabling precise control of the 3HV ratio in PHBV. <em>H. bluephagenesis</em> GZ05 was grown to 100 g L<sup>−1</sup> cell dry weight (CDW) containing 73 wt% PHBV consisting of 1.6 mol% 3HV in a 5000 L bioreactor. Thermal analysis demonstrated enhanced flexibility with higher 3HV content in PHBV.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 91-100"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145621007","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 a broad-spectrum glucose oxidase via substrate channel and linker design for enhanced lignocellulose bioconversion 通过底物通道和连接体设计设计广谱葡萄糖氧化酶以增强木质纤维素的生物转化

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2025-12-29 DOI: 10.1016/j.synbio.2025.11.016

Yong Feng , Xihua Chen , Zeyang Li , Zhong Ni , Zhengfen Wu , Zhongjian Guo , Fubao Sun , Huiqing Chen , Huayou Chen

{"title":"Engineering a broad-spectrum glucose oxidase via substrate channel and linker design for enhanced lignocellulose bioconversion","authors":"Yong Feng , Xihua Chen , Zeyang Li , Zhong Ni , Zhengfen Wu , Zhongjian Guo , Fubao Sun , Huiqing Chen , Huayou Chen","doi":"10.1016/j.synbio.2025.11.016","DOIUrl":"10.1016/j.synbio.2025.11.016","url":null,"abstract":"<div><div>Glucose oxidase (GOD) is a widely used enzyme in biotechnology, yet its narrow substrate specificity limits its application in complex bioconversion processes such as agricultural waste valorization. In this study, we employed synthetic biology and protein engineering strategies to engineer a broad-spectrum glucose oxidase from <em>Aureobasidium</em> sp. (AreGOD). Initially, site-directed mutagenesis at N82, a key gatekeeper at the dimer interface, modulated substrate channel geometry, leading to increased catalytic activity towards various sugars, particularly stachyose and xylose. Furthermore, systematic linker engineering between the spore anchor protein CotG and AreGOD revealed that flexible linkers, particularly the (GGGGS)<sub>5</sub> repeat (LK3), dramatically expanded the enzyme's substrate spectrum towards various mono-, di-, and oligosaccharides. The optimized spore-displayed AreGOD (CotG-LK3-AreGOD) exhibited strong synergistic effects with cellulase in wheat straw degradation, significantly enhancing the hydrolysis of cellulose, hemicellulose, and lignin. Our work demonstrates an effective and generalizable strategy for engineering substrate-promiscuous oxidases, highlighting the potential of integrative enzyme design for sustainable bioprocessing and agricultural biotechnology.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 218-228"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145883634","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

Enhanced AAV production via rational design of a novel pHelper vector integrated with HSV-1 helper genes 通过合理设计整合HSV-1辅助基因的新型辅助载体提高AAV的产生

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2026-01-15 DOI: 10.1016/j.synbio.2025.12.018

He Ren , Jianqi Nie , Zichuan Song , Wanting Mo , Yankun Yang , Zhonghu Bai

{"title":"Enhanced AAV production via rational design of a novel pHelper vector integrated with HSV-1 helper genes","authors":"He Ren , Jianqi Nie , Zichuan Song , Wanting Mo , Yankun Yang , Zhonghu Bai","doi":"10.1016/j.synbio.2025.12.018","DOIUrl":"10.1016/j.synbio.2025.12.018","url":null,"abstract":"<div><div>Adeno-associated virus (AAV) vectors are widely used in gene therapy owing to their safety, stability, and broad tissue tropism. However, current plasmid-based AAV manufacturing platforms suffer from low yield and high manufacturing cost, limiting their scalability for clinical and commercial applications. Rational engineering of pHelper vector offers an effective strategy to enhance AAV production. In this study, we engineered a novel helper vector (UL12-ICP22-miniHelper) by integrating <em>UL12</em> and <em>ICP22</em> genes from herpes simplex virus type 1 (HSV-1) into a size-reduced pHelper backbone (mini-pHelper) with partial deletion of <em>E2a</em> and <em>E4</em> regions. In a triple-plasmid transfection system, UL12-ICP22-miniHelper increased AAV5 vector yield from 1.35 × 10<sup>11</sup> to 2.85 × 10<sup>11</sup> vg/mL (2.11-fold) without altering the proportion of full capsids. Enhanced productivity was also observed across multiple serotypes, with increases of 2.24-fold for AAV1, 1.54-fold for AAV2, 1.88-fold for AAV6, and 2.03-fold for AAV9, while maintaining transduction efficiency. Mechanistic analysis indicated that the improved productivity was associated with elevated viral genome replication and increased Rep/Cap protein expression. Collectively, these results demonstrate that the novel UL12-ICP22-miniHelper provides a broadly applicable and cost-effective strategy for improving AAV vector manufacturing in both clinical and industrial applications.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 352-363"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145976680","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 amplification of NADPH-regenerating modules enhances cytidine biosynthesis in Escherichia coli nadph再生模块的整合扩增增强了大肠杆菌胞苷的生物合成

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2026-06-01 Epub Date: 2026-01-10 DOI: 10.1016/j.synbio.2025.12.009

Lu Liu , Xiangjun Zhang , Tengteng Zhu , Tong Ye , Zongqian Li , Wei Ding , Huiyan Liu , Haitian Fang

{"title":"Integrated amplification of NADPH-regenerating modules enhances cytidine biosynthesis in Escherichia coli","authors":"Lu Liu , Xiangjun Zhang , Tengteng Zhu , Tong Ye , Zongqian Li , Wei Ding , Huiyan Liu , Haitian Fang","doi":"10.1016/j.synbio.2025.12.009","DOIUrl":"10.1016/j.synbio.2025.12.009","url":null,"abstract":"<div><div>In <em>Escherichia coli</em>, cofactor imbalance serves as a crucial limiting factor in cytidine biosynthesis, with nicotinamide adenine dinucleotide phosphate (NADPH) insufficiency representing the principal metabolic barrier. To overcome this limitation, an integrated engineering strategy targeting the enhancement of NADPH metabolism was implemented. Via CRISPR-Cas9-mediated multiplex genomic editing and strong constitutive promoter replacement, three NADPH-regenerating modules were concurrently enhanced: the membrane-bound transhydrogenase (<em>pntAB</em>), the oxidative pentose phosphate pathway (<em>zwf</em>-encoded glucose-6-phosphate dehydrogenase), and the decarboxylation shunt (<em>gnd</em>-encoded 6-phosphogluconate dehydrogenase). After 54-hour fermentation in 500 mL shake flasks, the cytidine titer of the engineered strain NXBG-20 reached 7.83 g/L, representing a 9.10-fold increase compared to the start strain. Systematic multi-omics profiling revealed that the metabolic network had undergone substantial alterations. These alterations were characterized by the redirection of glycolytic flux towards nucleotide precursor substances and the enhancement of ribose-5-phosphate biosynthesis. This engineering approach not only establishes a novel microbial platform for cytidine bioproduction but also provides mechanistic insights into cofactor-driven metabolic flux control.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"12 ","pages":"Pages 320-329"},"PeriodicalIF":4.4,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145938532","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