Synthetic and Systems Biotechnology最新文献_第10页

MicroDFBEST: A dCas12b-derived dual-function base editor with programmable editing characteristics for microbial genetic engineering MicroDFBEST：一种dcas12b衍生的双功能碱基编辑器，具有可编程编辑特性，用于微生物基因工程

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2026-03-01 Epub Date: 2025-09-12 DOI: 10.1016/j.synbio.2025.09.013

Wen-Liang Hao , De-Zhi Geng , Yu-Feng Liu , Lai-Chuang Han , Zhe-Min Zhou , Wen-Jing Cui

Base editors (BEs) enable precise genome editing, but their use in microbes remains limited by restricted mutagenesis capabilities and narrow editing windows. Here, we reported MicroDFBEST, a novel dual-function base editor (DFBE) for microbes, by fusing the high-activity deaminases evoCDA1 and TadA9 with nuclease-deficient Cas12b from Bacillus hisashii (dBhCas12b). This engineered system enables simultaneous C-to-T and A-to-G editing within a 26–33 nt window, the broadest range reported for microbial DFBEs. The editing characteristics of MicroDFBEST can be easily adjusted by changing fusion protein expression and editing generations to create diverse mutant libraries. We show that the MicroDFBEST system enables both flexible gene expression modulation via random promoter (P_ylbP) diversification and targeted protein evolution through mutational hotspot scanning in native genomic contexts. This study offers a versatile platform enabling in situ gene regulation (e.g., biosynthetic gene clusters activation) and protein evolution (e.g., chassis optimization), with broad synthetic biology utility.

碱基编辑器（BEs）能够实现精确的基因组编辑，但它们在微生物中的应用仍然受到有限的诱变能力和狭窄的编辑窗口的限制。在这里，我们报道了MicroDFBEST，一种新的微生物双功能碱基编辑器（DFBE），通过将高活性脱氨酶evoCDA1和TadA9与来自hisashii芽孢杆菌的核酸酶缺陷Cas12b （dBhCas12b）融合。该工程系统能够在26 - 33nt的窗口内同时进行C-to-T和a -to- g编辑，这是目前报道的微生物DFBEs中范围最广的。通过改变融合蛋白表达和编辑代数，可以轻松调整MicroDFBEST的编辑特性，从而创建不同的突变文库。我们发现MicroDFBEST系统既可以通过随机启动子（PylbP）多样化灵活地调节基因表达，也可以通过突变热点扫描在原生基因组环境中实现靶向蛋白进化。该研究为原位基因调控（如生物合成基因簇激活）和蛋白质进化（如底盘优化）提供了一个多功能平台，具有广泛的合成生物学实用性。

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引用次数: 0

A near-infrared ratio fluorescent probe achieves mitochondrial hydrogen polysulfide imaging for monitoring ferroptosis in arthritis 近红外比例荧光探针实现线粒体多硫化氢成像监测关节炎中的铁下垂

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2026-03-01 Epub Date: 2025-09-02 DOI: 10.1016/j.synbio.2025.08.016

Ting Cao , Ziwen Xiao , Wenhua Dong , Hong Ma , Deyan Gong , Zhefeng Fan

Reactive sulfur (RSS) is a type of sulfur-containing molecule widely present in biological systems. Hydrogen polysulfide (H₂S_n, n > 1), as a member of the active sulfur family, plays an indispensable role in many physiological and pathological processes. Ferroptosis is a special cell death mode driven by iron-dependent lipid peroxidation, which is involved in the occurrence and development of various human diseases. Ferroptosis is manifested by increased lipid peroxidation and elevated levels of reactive oxygen species (ROS), which further lead to an increase in H₂S_n content in cells. Emerging research suggests a close association between ferroptosis and arthritis related diseases. This work successfully constructed a mitochondrial-targeted ratiometric near-infrared fluorescent probe for the specific detection of H₂S_n. The experimental results show that the probe (Cy-S₄) has a large Stokes shift (∼218 nm), excellent optical properties, extremely fast response time (8 s), high sensitivity (DL = 0.23 μM), and strong specificity. This probe has been successfully applied to tracking the content of H₂S_n in ferroptosis process and fluorescence imaging of H₂S_n in inflammatory cell mitochondria. Pathological section data confirmed that the probe has good in vivo imaging ability, and more importantly, in vivo arthritis imaging experiments showed that the expression of H₂S_n plays an important role in ferroptosis. These experimental results will provide a reliable monitoring tool for the treatment and prevention of arthritis, enriching the theoretical research related to this disease.

活性硫（RSS）是一类广泛存在于生物系统中的含硫分子。多硫化氢（H2Sn, n > 1）作为活性硫家族的一员，在许多生理病理过程中起着不可缺少的作用。铁下垂是一种由铁依赖性脂质过氧化作用驱动的特殊细胞死亡模式，参与了人类多种疾病的发生和发展。铁下垂表现为脂质过氧化和活性氧（ROS）水平升高，这进一步导致细胞中H2Sn含量增加。新兴研究表明，铁下垂与关节炎相关疾病密切相关。本工作成功构建了一种线粒体靶向比例荧光探针，用于特异检测H2Sn。实验结果表明，该探针（Cy-S4）具有较大的Stokes位移（~ 218 nm）、优异的光学性能、极快的响应时间（8 s）、高灵敏度（DL = 0.23 μM）和强特异性。该探针已成功应用于铁下垂过程中H2Sn含量的跟踪和炎症细胞线粒体中H2Sn的荧光成像。病理切片数据证实该探针具有良好的体内成像能力，更重要的是，体内关节炎成像实验表明H2Sn的表达在铁下垂中起重要作用。这些实验结果将为关节炎的治疗和预防提供可靠的监测工具，丰富该疾病的相关理论研究。

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引用次数: 0

Developing the protein quality control system in Streptomyces for selective translation of ultra-long full-length polyketide synthase mRNAs to enhance biosynthesis efficiency 建立链霉菌超长全长聚酮合成酶mrna选择性翻译的蛋白质量控制系统，提高生物合成效率

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2026-03-01 Epub Date: 2025-10-15 DOI: 10.1016/j.synbio.2025.10.004

Chanjuan Jiang , Kexin Hao , Shaoqian Wang , Ruofei Guo , Xiaochen Li , Lin Lv , Ji Luan , Hailong Wang

Genes that encode type I polyketide synthases (PKSs), which synthesize an array of invaluable drugs, typically exceed 10 kilobases in length. Truncated messenger RNAs (mRNAs) have been demonstrated to constitute the majority of PKS mRNAs; However, a method for the selective translation of full-length PKS mRNAs has yet to be established. Here we developed a protein quality control system in Streptomyces to selectively translate ultra-long full-length PKS mRNAs for enhanced biosynthesis efficiency. The Streptomyces protein quality control (strProQC) system comprises the switch RNA, which encapsulates the start codon and the ribosome binding site within a secondary structure, and the trigger RNA, which is the complementary counterpart to the switch RNA. The trigger RNA is placed at the 3′ end of the mRNA so that it can hybridize with the switch RNA to expose the translation-initiation region to ribosomes, enabling translation initiation. Truncated mRNAs do not contain the cis-trigger at their 3′ ends, therefore, their translation cannot be initiated and full-length mRNAs are selectively translated. We initially identified strong terminators and switch-trigger pairs to establish the strProQC system. Then the switch sequences in the strProQC system were further optimized by exchanging ribosome binding sites to improve the ON state strength by 2.8 folds and the ON/OFF ratio by 31.6 folds. Finally, the refined strProQC system was utilized for the selective translation of the full-length mRNAs of the 7.8-kb spinosad PKS gene (spnA) and the 25.7-kb rapamycin PKS gene (rapA). This resulted in a 1.4-fold increase in spinosad yields and a 4.7-fold increase in rapamycin yields, respectively, when compared to the yields obtained from the nonselective translation of both full-length and truncated PKS mRNAs.

编码I型聚酮合成酶（pks）的基因长度通常超过10千个碱基，pks可以合成一系列无价的药物。截断的信使rna （mrna）已被证明构成了大多数PKS mrna；然而，一种选择性翻译全长PKS mrna的方法尚未建立。我们在链霉菌中开发了一种蛋白质质量控制系统，以选择性地翻译超长全长PKS mrna，以提高生物合成效率。链霉菌蛋白质量控制（strProQC）系统包括将起始密码子和核糖体结合位点封装在二级结构中的开关RNA和与开关RNA互补的触发RNA。触发RNA被放置在mRNA的3 '端，这样它就可以与开关RNA杂交，将翻译起始区暴露给核糖体，从而实现翻译起始。截断的mrna在其3 '端不包含顺式触发器，因此，它们的翻译不能启动，全长mrna被选择性翻译。我们首先确定了强终止子和开关触发对来建立strProQC系统。然后通过交换核糖体结合位点进一步优化strProQC系统中的开关序列，使其ON状态强度提高2.8倍，ON/OFF比提高31.6倍。最后，利用改进的strProQC系统对7.8 kb的spinosad PKS基因（spnA）和25.7 kb的rapamycin PKS基因（rapA）的全长mrna进行选择性翻译。与全长和截断的PKS mrna的非选择性翻译相比，这导致spinosad产量增加1.4倍，雷帕霉素产量增加4.7倍。

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引用次数: 0

From consultors to collaborators – An SOP for advancing ethics engagement in science 从顾问到合作者——促进科学伦理参与的标准操作程序

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-12-01 Epub Date: 2025-06-23 DOI: 10.1016/j.synbio.2025.06.006

Varsha Aravind Paleri, Kristien Hens

Synthetic Biology (SynBio) is an interdisciplinary field that tackles global challenges in healthcare, sustainability, food production, among others. However, it also presents critical ethical concerns, including biosecurity, justice, and the unintended consequences of research biases. Addressing these concerns requires timely and systematic ethical evaluation. Conventional approaches often relegate ethics to an external, procedural role rather than embedding it within the core of scientific inquiry, leading to delayed engagement and associated challenges. This article introduces a structured, stage-wise Standard Operating Procedure (SOP) to support the systematic integration of ethics into scientific research. Particularly beneficial for early-career researchers and ethicists, this framework offers a practical method for incorporating ethical reflection throughout the research process.

合成生物学（SynBio）是一个跨学科领域，解决医疗保健、可持续性、食品生产等方面的全球挑战。然而，它也提出了关键的伦理问题，包括生物安全、正义和研究偏见的意外后果。解决这些问题需要及时和系统的伦理评估。传统的方法往往将伦理降级为外部的、程序性的角色，而不是将其嵌入科学探究的核心，从而导致参与的延迟和相关的挑战。本文介绍了一个结构化的、分阶段的标准操作程序（SOP），以支持伦理与科学研究的系统整合。该框架为在整个研究过程中纳入伦理反思提供了一种实用的方法，对早期职业研究人员和伦理学家尤其有益。

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 : 2025-12-01 Epub 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

Transcriptomic studies on the product stress response revealed that YCF1 is a beneficial factor for progesterone production in Yarrowia lipolytica 对产物胁迫反应的转录组学研究表明，YCF1是脂耶氏菌产生孕酮的有益因子

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-12-01 Epub Date: 2025-04-17 DOI: 10.1016/j.synbio.2025.04.008

Ying Wang , Ruosi Zhang , Mingdong Yao , Wenhai Xiao , Ying Wang , Ying-Jin Yuan

Progesterone is a widely used therapeutic hormone and a common precursor for the synthesis of pharmaceutical steroids in both mammals and plants. It has been successfully produced in heterologous microorganisms, but the market demand has not been met. The low progesterone yield, possibly due to product stress, may put pressure on the growth of strains and limit product synthesis efficiency. In this study, key pathways and genes that cause changes in amino acid and lipid metabolism and protein transport were identified through omics analysis. The expression of transporters leads to increased progesterone production and alleviated growth inhibition. Two related genes (gene IDs: 2912325 and 2908366) encoding the transporters glpF and SNQ2 improved production by 29.2 % and 51.7 %, respectively. Isoenzymes of native and exogenous transporters were screened and overexpressed. YCF1 from Saccharomyces cerevisiae exhibited the greatest benefit, increasing progesterone synthesis by 69.6 %. Our findings help reveal the impact of product stress on cellular metabolism and processes, providing research directions and literature support for the synthesis of other products.

黄体酮是一种广泛使用的治疗激素，也是哺乳动物和植物合成药物类固醇的常见前体。它已成功地在异源微生物中生产，但尚未满足市场需求。黄体酮产率低，可能是由于产物应激造成的，可能对菌株的生长造成压力，限制产物的合成效率。本研究通过组学分析，确定了引起氨基酸、脂质代谢和蛋白质转运变化的关键途径和基因。转运蛋白的表达增加了孕酮的产生，减轻了生长抑制。编码转运体glpF和SNQ2的两个相关基因（基因id: 2912325和2908366）分别提高了29.2%和51.7%的产量。筛选原生和外源转运体的同工酶并过表达。从酿酒酵母中提取的YCF1获益最大，使黄体酮的合成增加了69.6%。我们的发现有助于揭示产物应激对细胞代谢和过程的影响，为其他产物的合成提供研究方向和文献支持。

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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 : 2025-12-01 Epub 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 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 : 2025-12-01 Epub 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

Recombinant production of amaranthin and other betalain variants with yeast cell factories 用酵母细胞工厂重组生产苋菜素和其他甜菜素变体

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-12-01 Epub Date: 2025-05-21 DOI: 10.1016/j.synbio.2025.05.008

Christiane Glitz, Jane Dannow Dyekjær, Gian Maria Cristian Solimando, Paulo Marcelo Avila Neto, Daniela Rago, Mahsa Babaei, Irina Borodina

Betalains are a class of natural pigments found in plants of the Caryophyllales order. Betanin is the dominant betalain on the food colour market, even though over 80 other variants are known. Recombinant production of betanin has recently gained interest as a cost-efficient and sustainable alternative to traditional plant extraction, but the production of other betalain variants remains largely unexplored. We selected three glucuronosyltransferases from Amaranthus hypochondriacus, Chenopodium quinoa and Celosia argentea var. cristata and screened the enzymes in vivo in betanin-producing Saccharomyces cerevisiae. Instead of producing amaranthin, two of the enzymes led to the formation of the betalain bougainvillein-rI (betanidin 5-O-β-sophoroside). When expressed together with a UDP-dehydrogenase that allowed the synthesis of UDP-glucuronic acid, each enzyme converted betanin to amaranthin. Integration of the glucuronosyltransferases in a Yarrowia lipolytica betanin producer strain directly resulted in amaranthin production. In fed-batch fermentation, 2.97 g/L ± 29.3 mg/L of amaranthin was produced. Co-expression of the glucuronosyltransferases with a malonyltransferase from Hylocereus polyrhizus led to the formation of 6′-O-malonyl-amaranthin in S. cerevisiae and Y. lipolytica. This study expands the portfolio of natural food colourants that can efficiently be produced through microbial fermentation and contributes to elucidating the biosynthesis pathway of betalains.

甜菜素是在石竹目植物中发现的一类天然色素。甜菜素是食品色素市场上的主要甜菜素，尽管已知有80多种其他变体。重组甜菜素的生产作为一种具有成本效益和可持续性的传统植物提取替代品最近引起了人们的兴趣，但其他甜菜素变体的生产在很大程度上仍未被探索。从苋属、藜麦属和鸡冠花属中筛选出3种葡萄糖醛酸转移酶，并对产甜菜素的酿酒酵母进行了体内筛选。其中两种酶并没有产生紫红素，而是形成了甜菜素九重葛苷- ri（甜菜素5- 0 -β-槐苷）。当与允许合成udp -葡萄糖醛酸的udp -脱氢酶一起表达时，每种酶都将甜菜素转化为苋菜素。葡萄糖醛酸转移酶在聚脂耶氏菌β素产生菌中的整合直接导致了苋菜素的产生。分批补料发酵产生的苋菜素为2.97 g/L±29.3 mg/L。葡萄糖醛酸转移酶与多根Hylocereus malonyl转移酶的共同表达导致酿酒葡萄球菌和脂肪瘤葡萄球菌形成6′- o-丙二酰苋菜素。本研究扩大了微生物发酵生产天然食用色素的组合，有助于阐明甜菜素的生物合成途径。

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引用次数: 0

Metabolic engineering of Escherichia coli for squalene overproduction 过量生产角鲨烯的大肠杆菌代谢工程

IF 4.4 2区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Synthetic and Systems Biotechnology

Pub Date : 2025-12-01 Epub Date: 2025-06-07 DOI: 10.1016/j.synbio.2025.06.003

Jiangming Zhu , Yaping Mao , Hongchun Mo , Xuehui Dai , Yuhan Wu , Guangyi Wang , Zhanguang Feng , Ruirui Yue , Dongzhi Wei , Haili Liu , Yong Wang

Squalene, a lipophilic triterpene with multifaceted bioactivities, faces bioproduction bottlenecks in microbial hosts due to inefficient biosynthetic pathways and limited storage capacity. Here, we address these challenges through systems metabolic engineering integrating redox-balanced 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGR) variants and membrane lipid remodeling. By developing a hybrid HMGRs system combining NADPH-dependent and NADH-preferred enzymes, squalene production reached 852.06 ± 28.95 mg/L with balanced cofactor utilization. Subsequent engineering of membrane morphology and lipid metabolism generated lipid-enriched elongated cells, through the overexpression of dgs, murG and plsC, boosting squalene production to 970.86 ± 55.67 mg/L. Implementation of delayed induction strategies coupled with 10 % dodecane overlay as an in situ recovery system achieved a final squalene titer of 1267.01 mg/L in a 3 L bioreactor. Mechanistic studies revealed fatty acid (FA) and phosphatidylethanolamine (PE) as key reservoirs for squalene in E. coli, with dgs overexpression specifically promoting cellular elongation. This article provides comprehensive insights into engineering strategies and mechanistic perspectives, establishing a universal framework for hydrophobic metabolite biomanufacturing in prokaryotic hosts.

角鲨烯是一种具有多方面生物活性的亲脂性三萜，由于生物合成途径低效和储存能力有限，在微生物宿主中面临生物生产瓶颈。在这里，我们通过整合氧化还原平衡的3-羟基-3-甲基戊二酰辅酶A还原酶（HMGR）变体和膜脂重塑的系统代谢工程来解决这些挑战。通过建立nadph依赖酶和nadh优先酶的混合hmgr体系，角鲨烯产量达到852.06±28.95 mg/L，辅因子利用平衡。随后对膜形态和脂质代谢进行工程改造，通过过表达dgs、murG和plsC，生成富含脂质的细长细胞，使角鲨烯产量提高到970.86±55.67 mg/L。采用延迟诱导策略，加上10%十二烷覆盖作为原位回收系统，在3l生物反应器中最终获得了1267.01 mg/L的角鲨烯滴度。机制研究表明脂肪酸（FA）和磷脂酰乙醇胺（PE）是大肠杆菌中角鲨烯的关键储存库，dgs过表达特异性地促进细胞伸长。本文从工程策略和机理角度提供了全面的见解，建立了原核宿主中疏水代谢物生物制造的通用框架。

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