Synthetic and Systems Biotechnology最新文献_第10页

Malic enzyme-based system for transhydrogenation between nicotinamide cofactors 基于苹果酶的烟酰胺辅助因子间转氢化系统

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-21 DOI: 10.1016/j.synbio.2025.08.009

Haizhao Xue , Yinghan Hu , Aabid Manzoor Shah , Xueying Wang , Xiaojia Guo , Yanzhe Huang , Zongbao K. Zhao

The nicotinamide cofactors including nicotinamide adenine dinucleotide (NAD) and its phosphate (NADP) play important roles in facilitating redox reactions for energy metabolism and biosynthesis. To expand the cofactor menu, a non-natural cofactor nicotinamide cytosine dinucleotide (NCD) has been introduced recently. The reduced forms of these cofactors carry reducing equivalents that are essential for cellular metabolism. However, there is a long standing challenge to rationally transfer reducing equivalent from one cofactor to another, albeit such process is highly demanding in metabolic engineering. This study develops a new approach based on malic enzyme (ME)-mediated transhydrogenation to enable reducing equivalents exchange among different cofactors. We used wild-type ME, MaeB and an engineered ME∗ that favors NAD, NADP and NCD, respectively, to demonstrate such conversions. When an in vitro system initiated with equal amount of NADH and NCD in the presence of ME, ME∗ and excess amount of pyruvate was held for 2 h, up to 65 % NADH was consumed and 57 % NCDH was generated. When implemented into NCD self-sufficient Escherichia coli cells, the system directed reducing equivalents toward NCDH-linked formation of lactate. Overall, this work offers an effective strategy to regulate intracellular reducing equivalents that may serve as a novel tool for metabolic engineering and synthetic biology.

烟酰胺辅助因子包括烟酰胺腺嘌呤二核苷酸（NAD）及其磷酸（NADP），在促进能量代谢和生物合成的氧化还原反应中起重要作用。为了扩大辅助因子的范围，最近引入了一种非天然的辅助因子烟酰胺胞嘧啶二核苷酸（NCD）。这些辅因子的还原形式携带细胞代谢所必需的还原等价物。然而，尽管在代谢工程中这一过程要求很高，但如何合理地将还原当量从一种辅因子转移到另一种辅因子仍是一个长期存在的挑战。本研究开发了一种基于苹果酸酶（ME）介导的转氢反应的新方法，以减少不同辅因子之间的等价物交换。我们分别使用野生型ME、MaeB和有利于NAD、NADP和NCD的工程ME *来演示这种转化。在体外系统中，在ME、ME *和过量的丙酮酸存在下，用等量的NADH和NCD启动2小时，高达65%的NADH被消耗，57%的NCDH被生成。当将其应用于非传染性疾病自给自足的大肠杆菌细胞时，该系统将还原等量物导向与非传染性疾病相关的乳酸形成。总的来说，这项工作提供了一种有效的策略来调节细胞内还原当量，可能作为代谢工程和合成生物学的新工具。

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引用次数: 0

De novo biosynthesis of Asperosaponin VI in Saccharomyces cerevisiae

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-19 DOI: 10.1016/j.synbio.2025.08.007

Lin Hao , Guiru Dong , Tianzhen Sun , Jingyan Liu , Hui Wu , Fahui Li , Weiguo Song , Xiaozhou Luo , Jian Zhang , Yanan Qiao

Asperosaponin VI (ASA VI), the primary bioactive triterpenoid saponin marker of Dipsacus asper Wall. (Chinese Pharmacopoeia 2020), possesses significant neuroprotective, anti-inflammatory, and osteogenic activities. However, its low natural abundance limits large-scale production. In this study, we reported the first complete biosynthetic reconstruction of ASA VI in Saccharomyces cerevisiae using a modular synthetic biology strategy. The pathway included in situ UDP-arabinose (UDP-Ara) biosynthesis via heterologous expression of AtUGDH3, GuUXS2, and GuUXE1; triterpenoid scaffold generation through ERG9, ERG1, CqBAS1, and CqCYP716A78 for oleanolic acid (OA) production; and downstream modifications including C-23 hydroxylation by multicopy-expressed CaCYP714E19, stepwise glucosylation at C-28 by CaUGT73AD1 and CaUGT73C8, and C-3 arabinosylation by AsUGT99D1 to yield ASA VI. LC-MS analysis confirmed ASA VI biosynthesis and the accumulation of key intermediates (OA, HED, HED-28-Glc, and HED-28-Glc-Glc). Although production remained at trace levels (395 ng/L), pathway analysis suggested that the downstream glycosylation steps and UDP-Ara supply could be the major rate-limiting factors. This work established a microbial chassis for the sustainable synthesis of ASA VI and related arabinosylated saponins.

Asperosaponin VI （ASA VI）是一种主要的生物活性三萜皂苷标记物。（中国药典2020），具有显著的神经保护、抗炎和成骨活性。然而，其天然丰度低限制了大规模生产。在这项研究中，我们报道了首次使用模块化合成生物学策略在酿酒酵母中完成ASA VI的生物合成重建。该途径包括通过异源表达AtUGDH3、GuUXS2和GuUXE1原位合成udp -阿拉伯糖（UDP-Ara）；通过ERG9、ERG1、CqBAS1和CqCYP716A78生成三萜支架，用于齐墩果酸（OA）的生产；下游修饰包括C-23被多拷贝表达的CaCYP714E19羟基化，C-28被CaUGT73AD1和CaUGT73C8逐步糖基化，C-3被AsUGT99D1阿拉伯糖基化，从而产生ASA VI。LC-MS分析证实了ASA VI的生物合成和关键中间体（OA、HED、HED-28- glc和HED-28- glc - glc）的积累。虽然产量保持在微量水平（395 ng/L），但途径分析表明，下游糖基化步骤和UDP-Ara供应可能是主要的限速因素。这项工作为ASA VI和相关的阿拉伯糖基化皂苷的可持续合成建立了一个微生物基础。

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引用次数: 0

Genome mining and characterization of a heme-dependent enzyme catalyzing intermolecular Nitrogen–Nitrogen bond formation in hydrazinosuccinic acid biosynthesis 一种在肼琥珀酸生物合成中催化分子间氮-氮键形成的血红素依赖酶的基因组挖掘和表征

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-18 DOI: 10.1016/j.synbio.2025.08.006

Jingkun Shi , Zhijie Zhao , Jian Yang , Ziyang Cheng , Hu Li , Yu Liu , Guiyun Zhao , Miaolian Wu , Yi-Ling Du

Nitrogen–nitrogen (N–N) bond-forming enzymes are rare but play vital roles in both primary and secondary metabolism. Guided by a nitric oxide synthase (NOS)-based genome mining strategy, we report the discovery and characterization of a new heme-dependent enzyme system that catalyzes intermolecular N–N bond formation. Using both in vivo and in vitro reconstitution approaches, we demonstrated that a protein complex, comprising a heme enzyme and a 2[4Fe–4S] ferredoxin partner, mediates the coupling of the α-amine group of l-aspartate with inorganic nitrogen oxide species, such as nitrite or nitric oxide, to generate hydrazinosuccinic acid, a key biosynthetic precursor in several natural product pathways. Structural modeling and site-directed mutagenesis suggest a plausible catalytic mechanism involving the formation of a reactive nitrogen intermediate, potentially a heme-bound nitrene species. These findings reveal a new family of N–N bond-forming biocatalysts that leverage inorganic nitrogen sources, offering valuable tools for genome mining and the synthetic biology.

氮-氮（N-N）键形成酶是罕见的，但在初级和次级代谢中都起着重要作用。在基于一氧化氮合酶（NOS）的基因组挖掘策略的指导下，我们报告了一种催化分子间N-N键形成的新的血红素依赖酶系统的发现和表征。利用体内和体外重构方法，我们证明了一种蛋白质复合物，包括血红素酶和2[4Fe-4S]铁氧还蛋白伴侣，介导l-天冬氨酸α-胺基与无机氮氧化物（如亚硝酸盐或一氧化氮）的偶联，生成肼琥珀酸，这是几种天然产物途径中的关键生物合成前体。结构建模和位点定向诱变提示了一种可能的催化机制，涉及活性氮中间体的形成，可能是血红素结合的亚硝基物质。这些发现揭示了一个利用无机氮源形成N-N键的新生物催化剂家族，为基因组挖掘和合成生物学提供了有价值的工具。

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引用次数: 0

Harnessing synthetic biology for tetraterpenoid astaxanthin production: Recent advances and challenges 利用合成生物学生产四萜类虾青素：最新进展和挑战

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-18 DOI: 10.1016/j.synbio.2025.08.005

Yue Hou , Ailin Guan , Xuefen Fan , Jiufu Qin

Astaxanthin, a potent lipid-soluble ketocarotenoid, exists in various stereoisomeric, geometric isomeric, and esterified forms. Its unique molecular configuration and biological activity confer significant advantages for pharmaceutical applications and nutraceutical supplementation. While widely distributed in natural environments, particularly abundant in marine ecosystems, astaxanthin production via conventional methods fails to meet escalating market demands and consumer preference for natural products. Recent synthetic biology advances enable engineered microbial cell factories for astaxanthin production. Through precise genome editing and metabolic reprogramming, these systems offer a sustainable, efficient alternative to traditional synthesis of this high-value antioxidant. In this review, we highlight the latest advancements in constructing artificial cell factories for the production of astaxanthin. Specially, we systematically examine current breakthroughs in synthetic biology-enabled astaxanthin manufacturing, with emphasis on three dimensions: “point” (regulation of key enzymatic activity and expression levels), “line” (pathway-level spatial coordination to balance intermediate flux), and “plane” (system-level metabolic harmonization to address product toxicity and pathway-host metabolic incompatibility). We further discuss critical gaps requiring interdisciplinary innovation to realize the full potential of microbial cell factories in sustainable astaxanthin manufacturing production.

虾青素是一种有效的脂溶性类酮胡萝卜素，以各种立体异构体、几何异构体和酯化形式存在。其独特的分子结构和生物活性使其在制药和营养补充剂方面具有显著的优势。虽然虾青素广泛分布于自然环境中，特别是在海洋生态系统中含量丰富，但通过传统方法生产虾青素无法满足日益增长的市场需求和消费者对天然产品的偏好。合成生物学的最新进展使工程微生物细胞工厂能够生产虾青素。通过精确的基因组编辑和代谢重编程，这些系统为这种高价值抗氧化剂的传统合成提供了可持续、高效的替代方案。本文就虾青素人工细胞工厂的研究进展作一综述。特别地，我们系统地研究了合成生物学使虾青素制造的当前突破，重点是三个维度：“点”（关键酶活性和表达水平的调节），“线”（途径水平的空间协调以平衡中间通量）和“面”（系统水平的代谢协调以解决产品毒性和途径-宿主代谢不相容性）。我们进一步讨论了需要跨学科创新的关键差距，以实现微生物细胞工厂在可持续虾青素制造生产中的全部潜力。

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引用次数: 0

Engineering a drug-inducible pyroptosis platform enables precise tumor suppression in colorectal cancer 设计一个药物诱导的焦亡平台可以精确抑制结直肠癌的肿瘤

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-14 DOI: 10.1016/j.synbio.2025.08.004

Xiang Yao , Yu Wei , Yuan Gao , Lei Li , Junchi Liu , Wenmin Zhou , Tao Yan , Letian Gong , Yang Zhou , Ganglong Gao

Colorectal cancer remains a leading cause of cancer-related mortality, with long-term survival rates hindered by chemoresistance and an immunosuppressive tumor microenvironment. Gene-based therapies offer high specificity but are limited by challenges such as off-target effects, inefficient delivery, and systemic toxicity. Here, we report the design and functional validation of a chemically inducible gene circuit that harnesses Gasdermin E (GSDME) to trigger pyroptotic cell death on demand. We substituted its native proteolytic activation motif with a customized protease recognition sequence. By engineering inducible protease variants whose activity is tightly regulated by an orally bioavailable, clinically approved small molecule, we achieved precise temporal control of pyroptosis. In patient-derived organoid models, administration of the inducer led to rapid GSDME cleavage, pore formation, and robust cell lysis. In a xenograft model, oral treatment with the approved drug led to marked tumor growth inhibition. This strategy utilizes the safety and pharmacokinetics of an approved drug to enable programmable cell death, providing a versatile platform for the targeted elimination of treatment-resistant tumors.

结直肠癌仍然是癌症相关死亡的主要原因，其长期生存率受到化疗耐药和免疫抑制肿瘤微环境的阻碍。基于基因的治疗具有高特异性，但受到脱靶效应、低效递送和全身毒性等挑战的限制。在这里，我们报道了一种化学诱导基因电路的设计和功能验证，该电路利用Gasdermin E （GSDME）根据需要触发热亡细胞死亡。我们用定制的蛋白酶识别序列取代了其天然的蛋白水解激活基序。通过工程诱导的蛋白酶变体，其活性由口服生物可利用的临床批准的小分子严格调节，我们实现了对焦亡的精确时间控制。在患者来源的类器官模型中，使用诱导剂导致GSDME快速裂解、孔形成和稳健的细胞裂解。在异种移植物模型中，经批准的药物口服治疗可显著抑制肿瘤生长。该策略利用已批准药物的安全性和药代动力学来实现可编程细胞死亡，为靶向消除治疗耐药肿瘤提供了一个多功能平台。

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引用次数: 0

Engineering robust Saccharomyces cerevisiae cell factory for improving fatty alcohol biosynthesis 设计健壮的酿酒酵母细胞工厂以改善脂肪醇的生物合成

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-13 DOI: 10.1016/j.synbio.2025.08.003

Sijia Kong , Wei Yu , Zulin Wu , Ning Gao , Yongjin J. Zhou

Engineering yeast cell factories is a feasible approach to produce value chemicals from renewable feedstocks. However, during the production process, reprogramming of the internal metabolic pathways of yeast cells and environmental stress always compromises its production performance. Here, we engineered the robust Saccharomyces cerevisiae to enhance the production of fatty alcohols by downregulating the expression of target of rapamycin gene TOR1 and deleting histone deacetylase gene HDA1 in S. cerevisiae. The enhanced cellular robustness resulted in the extended chronological lifespan (CLS) through metabolic balance and stress response regulation, thus increasing the production of fatty alcohols by up to 56 %. This strategy may be used as a general strategy for building effective microbial cell factories.

工程酵母细胞工厂是利用可再生原料生产有价值化学品的可行途径。然而，在生产过程中，酵母细胞内部代谢途径的重编程和环境胁迫往往会影响其生产性能。在这里，我们设计了健壮的酿酒酵母，通过下调雷帕霉素靶基因TOR1的表达和删除酿酒酵母中组蛋白去乙酰化酶基因HDA1来提高脂肪醇的产生。增强的细胞健壮性通过代谢平衡和应激反应调节导致了按时间顺序寿命（CLS）的延长，从而使脂肪醇的产量增加了56%。该策略可作为建立有效微生物细胞工厂的一般策略。

引用次数: 0

Development and application of synthetic biology tools for improved production of human breast milk components 合成生物学工具的开发和应用，以改善人类母乳成分的生产

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-07 DOI: 10.1016/j.synbio.2025.08.002

Manxiang Zhu , Xianhao Xu , Xinyu Bi , Shixiu Cui , Yanfeng Liu , Guocheng Du , Xueqin Lv , Long Liu , Jianghua Li

Human breast milk (HBM) is composed of various components that are crucial for providing essential nutrients and enhancing infant immune system. Recently, the synthesis of HBM components through microbial fermentation has garnered significant attention due to its potential to reduce production costs, simplify manufacturing processes, and mitigate environmental pollution. However, this approach results in low yield and is difficult to scale up at the industrial level. Therefore, various synthetic biology tools have been used to enhance the efficiency of HBM component synthesis. This review first summarizes several synthetic biology tools that may improve HBM component production. Next, we have summarized HBM component production using microbial cell factories. Finally, we have summarized the challenges and opportunities presented by the construction of cell factories for the synthesis of HBM using synthetic biology tools. This article therefore provides a general guide to the construction of microbial cell factories for HBM components.

人类母乳（HBM）由各种成分组成，这些成分对提供必需的营养和增强婴儿免疫系统至关重要。最近，通过微生物发酵合成HBM组分因其降低生产成本、简化制造工艺和减轻环境污染的潜力而受到了极大的关注。然而，这种方法导致产量低，难以在工业水平上扩大规模。因此，各种合成生物学工具被用于提高HBM成分的合成效率。本文首先综述了几种可能提高HBM成分生产的合成生物学工具。接下来，我们总结了利用微生物细胞工厂生产HBM组件。最后，总结了利用合成生物学工具构建HBM合成细胞工厂所面临的挑战和机遇。因此，本文为HBM组分微生物细胞工厂的建设提供了一般指导。

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引用次数: 0

Engineered bacteriophage-based bioimaging Technology: Development and applications 基于工程噬菌体的生物成像技术：发展与应用

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-06 DOI: 10.1016/j.synbio.2025.07.009

Yuanzhao Shen , Lichang Sun , Jun Li , Xin Zhou , Ran Wang

Engineered bacteriophages (phages) have emerged as powerful and versatile tools for bioimaging, owing to their natural specificity for bacterial targets and their amenability to functional modification. This review summarizes recent advances in the development and application of phage-based imaging probes, with a particular focus on surface functionalization techniques and genetic engineering strategies used to construct functional phage imaging agents. These engineered phage probes have been applied across diverse imaging modalities, including fluorescence, magnetic resonance imaging (MRI), nuclear imaging, near-infrared (NIR) optical imaging, and surface-enhanced Raman scattering (SERS), etc. and have been utilized to enable highly sensitive detection of bacterial pathogens, improved diagnosis of infectious diseases, and monitoring of tissue engineering processes. Despite these innovations, critical challenges remain in ensuring robust target specificity, precise control of labeling stoichiometry, and favorable biocompatibility. Addressing issues such as non-specific probe binding, signal quenching, and immunogenicity will be crucial to fully realize the potential of phage-based bioimaging. Looking ahead, this review discusses future directions for next-generation phage imaging platforms with enhanced specificity, multiplexed functionality, and improved translational potential for clinical diagnostics.

工程噬菌体（噬菌体）由于其对细菌靶点的天然特异性和对功能修饰的适应性，已成为生物成像的强大而多功能的工具。本文综述了噬菌体成像探针的发展和应用的最新进展，重点介绍了表面功能化技术和用于构建功能性噬菌体成像剂的基因工程策略。这些工程噬菌体探针已被应用于多种成像方式，包括荧光、磁共振成像（MRI）、核成像、近红外（NIR）光学成像和表面增强拉曼散射（SERS）等，并已被用于实现细菌病原体的高灵敏度检测，改进传染病的诊断，以及组织工程过程的监测。尽管有这些创新，关键的挑战仍然是确保强大的目标特异性，精确控制标记化学计量，以及良好的生物相容性。解决诸如非特异性探针结合、信号猝灭和免疫原性等问题对于充分发挥噬菌体生物成像的潜力至关重要。展望未来，本综述讨论了下一代噬菌体成像平台的未来发展方向，这些平台具有增强的特异性、多路功能和改善的临床诊断转化潜力。

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引用次数: 0

Engineering non-P450 3-hydroxylase for de novo synthesizes catechol-containing compounds in Escherichia coli 工程非p450 3-羟化酶在大肠杆菌中重新合成含儿茶酚的化合物

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-05 DOI: 10.1016/j.synbio.2025.07.014

Xing-Run Zheng, Guan-Peng Li, Qian-Hui Chen, Jian-Zhong Liu

Catechols (such as l-DOPA, caffeic acid and hydroxytyrosol, etc.) are a class of phenolic derivatives with ortho-hydroxyl groups which represents various bioactivities including antioxidative, anti-inflammatory, antiviral, and anticancer properties. Non-P450-dependent 3′-hydroxylases HpaBC are the rate-limiting enzymes in catechol biosynthesis. Herein, different HpaB/HpaC combinations were first investigated. The best combinations of KpHpaB from Klebsiella pneumoniae and PaHpaC from Pseudomonas aeruginosa (or SeHpaC from Salmonella enterica) were obtained for the de novo synthesis of l-DOPA in E. coli, resulting in 1838.56 mg/L l-DOPA (or 1822.99 mg/L l-DOPA). The highest production of caffeic acid and hydroxytyrosol were obtained with the enzyme combinations of PaHpaB from P. aeruginosa and SeHpaC from S. enterica, and PlHpaB from Photorhabdus luminescens and KpHpaC from K. pneumoniae, respectively. Next, PaHpaB and PlHpaB were further engineered to improve their catalytic efficiency by the semi-rational method. PaHpaB^A211W and PlHpaB^S210G were obtained. The titer of caffeic acid was further increased to 1281.25 mg/L without l-DOPA accumulation using the PaHpaB^A211W-UTR-SeHpaC hybrid. The production of hydroxytyrosol was further enhanced to 1681.42 mg/L using the combination of PlHpaB^S210G-UTR- KpHpaC. The production of l-DOPA, caffeic acid and hydroxytyrosol was increased using these hybrids of HpaB/HpaC by 4.6-fold, 10.1-fold, and 8.4-fold compared to EcHpaBC from Escherichia coli, respectively. This work demonstrates that pairing of HpaB/HpaC and engineering HpaB is an powerful method for improving 3-hydroxylase activity and the production of catechol-containing compounds.

儿茶酚（如左旋多巴、咖啡酸和羟基酪醇等）是一类具有邻羟基的酚类衍生物，具有抗氧化、抗炎、抗病毒和抗癌等多种生物活性。非p450依赖性3 ' -羟化酶HpaBC是儿茶酚生物合成中的限速酶。本文首先研究了不同的HpaB/HpaC组合。肺炎克雷伯菌的KpHpaB和铜绿假单胞菌的PaHpaC（或肠沙门氏菌的SeHpaC）的最佳组合在大肠杆菌中重新合成L - dopa，得到的L - dopa为1838.56 mg/L（或1822.99 mg/L）。铜绿假单胞菌（P. aeruginosa）的PaHpaB和肠链球菌（S. enterica）的SeHpaC、发光光habus luminesens的PlHpaB和肺炎卡伯菌（K. pneumoniae）的KpHpaC组合酶的咖啡酸和羟基酪醇产量最高。接下来，对PaHpaB和PlHpaB进行半合理修饰，进一步提高其催化效率。得到了PaHpaBA211W和PlHpaBS210G。使用PaHpaBA211W-UTR-SeHpaC杂交，咖啡酸滴度进一步提高到1281.25 mg/L，没有L - dopa积累。结合PlHpaBS210G-UTR- KpHpaC，羟基酪醇的产量进一步提高到1681.42 mg/L。与大肠杆菌中的EcHpaBC相比，这些HpaB/HpaC杂交体的l-DOPA、咖啡酸和羟基酪醇的产量分别增加了4.6倍、10.1倍和8.4倍。这项工作表明，HpaB/HpaC与工程HpaB的配对是提高3-羟化酶活性和含儿茶酚化合物生产的有力方法。

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引用次数: 0

Engineered membraneless organelles in Corynebacterium glutamicum for enhanced indigoidine biosynthesis and antimicrobial peptide production 谷氨酸棒状杆菌的工程无膜细胞器增强靛蓝素生物合成和抗菌肽生产

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-08-05 DOI: 10.1016/j.synbio.2025.08.001

Manman Sun , Yimeng Zhao , Rodrigo Ledesma-Amaro , Jin Gao , Xiuxia Liu , Zhonghu Bai , Alex Xiong Gao , Peng Wang

Liquid-liquid phase separation (LLPS)-driven membraneless organelles (MLOs) have been employed to enhance metabolic efficiency in various microbial cell factories. However, their application in the industrial bacterium Corynebacterium glutamicum has not been explored. Here, we report the formation of liquid protein condensates in C. glutamicum using the RGG domain of Caenorhabditis elegans LAF-1. We optimized conditions for condensate formation, including the pre-induction period, inducer concentration, and cultivation temperature. Using the indigoidine biosynthesis pathway as a model, we demonstrated that LLPS-mediated MLOs enhanced indigoidine production. Furthermore, we applied these MLOs to modulate the toxicity of antimicrobial peptides (AMPs) to host cells, facilitating the expression of AMPs, including melittin and lactoferricin B. These findings provide insights into MLOs engineering in C. glutamicum and suggest broader applications of LLPS-mediated systems in industrial biotechnology.

液-液相分离驱动的无膜细胞器（MLOs）已被用于提高各种微生物细胞工厂的代谢效率。然而，它们在工业细菌谷氨酸棒状杆菌中的应用尚未探索。在这里，我们报道了利用秀丽隐杆线虫la1的RGG结构域在C. glutamicum中形成液体蛋白凝析物。我们优化了凝析液形成的条件，包括诱导前时间、诱导剂浓度和培养温度。以靛蓝苷生物合成途径为模型，我们证明了llps介导的MLOs促进了靛蓝苷的产生。此外，我们将这些MLOs应用于调节抗菌肽（AMPs）对宿主细胞的毒性，促进AMPs的表达，包括蜂毒蛋白和乳铁蛋白b。这些发现为谷氨酸梭菌的MLOs工程提供了见解，并为llps介导的系统在工业生物技术中的更广泛应用提供了建议。

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引用次数: 0