Metabolic engineering最新文献_第8页

Metabolic engineering of Corynebacterium glutamicum for increased cis, cis-muconate production from plant-derived p-hydroxycinnamates via deregulated pathway flux and increased CoA intermediate availability 谷氨酸棒状杆菌的代谢工程，通过解除调控的途径通量和增加辅酶a中间体的可用性，增加植物源的对羟基肉桂酸的顺式、顺式肉桂酸的产量。

IF 6.8 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

Pub Date : 2025-08-12 DOI: 10.1016/j.ymben.2025.08.004

Fabia Weiland, Kyoyoung Seo, Franka Janz, Marius Grad, Lea Geldmacher, Michael Kohlstedt, Judith Becker, Christoph Wittmann

Lignocellulosic biomass represents a promising renewable feedstock for sustainable biochemical production, with p-hydroxycinnamates emerging as key aromatic building blocks derived from agricultural residues and grassy plants. C. glutamicum has recently been engineered to produce cis, cis-muconate (MA), a high-value platform chemical used in biobased plastics, resins, and specialty chemicals. However, unlike other aromatics, the metabolism of the p-hydroxycinnamates p-coumarate, ferulate, and caffeate in MA-producing C. glutamicum is inefficient, limiting MA production performance. Here, we discovered that p-hydroxycinnamate metabolism, encoded by the phd operon, is repressed by the local repressor PhdR under glucose-rich conditions, while the global regulator GlxR activates the pathway in the absence of glucose. The deregulated C. glutamicum MA-10 lacking phdR exhibited an up to 98-fold increase in the conversion of p-coumarate, ferulate, and aromatic mixtures derived from plant waste into MA. Transcriptomic and metabolomic analyses revealed strong induction of the phd operon in strain MA-10 and a marked increase in intracellular aromatic CoA-esters and acetyl-CoA, indicating enhanced flux through the p-hydroxycinnamate degradation pathway. ¹³C-tracer studies demonstrated a substantial contribution of aromatic side-chain carbon to central metabolic pathways, supporting biomass formation and enabling MA production even in the absence of sugars. Additionally, MA-10 showed broadened substrate flexibility, degrading cinnamate into MA and methoxylated cinnamates into valuable benzoate derivatives. The strain also successfully converted aromatics from real straw lignin hydrolysates into MA. Our findings reveal the potential of targeted regulatory engineering to optimize C. glutamicum for lignin valorization. The newly developed strain MA-10 provides a robust platform for the biobased production of MA from lignocellulosic feedstocks, paving the way for sustainable and economically viable biorefinery processes.

木质纤维素生物质代表了一种有前途的可持续生化生产的可再生原料，对羟基肉桂酸酯作为从农业残留物和禾草植物中提取的关键芳香基石。C. glutamicum最近被设计用于生产cis， cis-muconate (MA)，这是一种高价值的平台化学品，用于生物基塑料，树脂和特种化学品。然而，与其他芳香烃不同的是，对羟基肉桂酸、对香豆酸、阿魏酸和咖啡酸在产生MA的C. glutamum中的代谢效率很低，限制了MA的生产性能。在这里，我们发现由phd操纵子编码的对羟基肉桂酸代谢在富含葡萄糖的条件下被局部抑制因子PhdR抑制，而全局调节因子GlxR在缺乏葡萄糖的情况下激活该途径。缺乏phdR的不受调控的谷氨酰胺MA-10在从植物废物中提取的对香豆酸盐、阿魏酸盐和芳香混合物转化为MA的能力增加了98倍。转录组学和代谢组学分析显示，菌株MA-10的phd操纵子被强烈诱导，细胞内芳香辅酶a酯和乙酰辅酶a显著增加，表明通过对羟基肉桂酸降解途径的通量增强。13c示踪研究表明芳香侧链碳对中心代谢途径的重要贡献，支持生物量的形成，即使在没有糖的情况下也能产生MA。此外，MA-10表现出更宽的底物柔韧性，将肉桂酸降解为MA，并将甲氧基化的肉桂酸转化为有价值的苯甲酸衍生物。该菌株还成功地将真正的秸秆木质素水解产物中的芳烃转化为MA。我们的研究结果揭示了靶向调控工程优化谷氨酰胺木质素增值的潜力。新开发的菌株MA-10为木质纤维素原料的生物基生产MA提供了一个强大的平台，为可持续和经济上可行的生物炼制工艺铺平了道路。

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引用次数: 0

Heterologous integration-assisted metabolic engineering in Escherichia coli for elevated D-pantothenic acid production 外源整合辅助大肠杆菌代谢工程提高d -泛酸产量。

IF 6.8 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

Pub Date : 2025-08-11 DOI: 10.1016/j.ymben.2025.08.003

Kuo Zhao , Hailin Gao , Mengnan Han , Bo Zhang , Zhiqiang Liu , Shuping Zou , Yuguo Zheng

D-pantothenic acid (D-PA) is a vital water-soluble vitamin with diverse industrial applications, driving the demand for efficient microbial production. Here, we rationally engineered an Escherichia coli strain to enhance D-PA production through metabolic engineering. First, to enhance carbon utilization efficiency, competing byproduct pathways were deleted and the pentose phosphate pathway was downregulated. Next, the glucose and β-alanine transport systems were strategically enhanced, and cofactor availability was improved through engineering NADPH regeneration and ATP recycling pathways. Subsequently, pathway engineering was applied to fine-tune the expression of heterologous enzymes, thereby enhancing the metabolic pull toward D-PA biosynthesis. To enhance the supply of one-carbon donor required by the rate-limiting enzyme ketopantoate hydroxymethyltransferase (KPHMT), a heterologous 5,10-methylenetetrahydrofolate biosynthesis module was introduced. Finally, dynamic regulation of isocitrate synthase and pantothenate kinase was implemented to balance cell growth and D-PA production. As a result of the integrated metabolic engineering strategies, the final strain DPZ28/P31 achieved a D-PA titer of 98.6 g/L and a yield of 0.44 g/g glucose in a two-stage fed-batch fermentation. These findings provide valuable insights for industrial-scale production of D-PA and related compounds.

d -泛酸（D-PA）是一种重要的水溶性维生素，具有多种工业应用，推动了对高效微生物生产的需求。本研究通过代谢工程对大肠杆菌菌株进行合理改造，提高D-PA的产量。首先，为了提高碳利用效率，删除竞争性副产物途径，下调戊糖磷酸途径。接下来，葡萄糖和β-丙氨酸运输系统被战略性地增强，通过工程NADPH再生和ATP循环途径提高辅助因子的可用性。随后，途径工程应用于微调异种酶的表达，从而增强对D-PA生物合成的代谢拉动。为了提高酮托酸羟甲基转移酶（KPHMT）所需的单碳供体的供应，引入了异源的5,10-亚甲基四氢叶酸生物合成模块。最后，通过动态调节异柠檬酸合成酶和泛酸激酶来平衡细胞生长和D-PA的产生。通过综合代谢工程策略，最终菌株DPZ28/P31在两段补料分批发酵中获得了D-PA滴度为98.6 g/L，葡萄糖产量为0.44 g/g。这些发现为D-PA及其相关化合物的工业规模生产提供了有价值的见解。

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引用次数: 0

Metabolic flux and flux balance analyses indicate the relevance of metabolic thermogenesis and aerobic glycolysis in cancer cells 代谢通量和通量平衡分析表明，代谢产热和有氧糖酵解在癌细胞中的相关性。

IF 6.8 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

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

Nobuyuki Okahashi , Tomoki Shima , Yuya Kondo , Chie Araki , Shuma Tsuji , Akane Sawai , Hikaru Uehara , Susumu Kohno , Hiroshi Shimizu , Chiaki Takahashi , Fumio Matsuda

Adenosine triphosphate (ATP) regeneration by substrate-level phosphorylation is a general feature of cancer metabolism, even under normoxic conditions (aerobic glycolysis). However, it is unclear why cancer cells prefer inefficient aerobic glycolysis over the highly efficient process of oxidative phosphorylation for ATP regeneration. To investigate the metabolic principles underlying aerobic glycolysis, we performed ¹³C-metabolic flux analysis of 12 cultured cancer cell lines and explored the metabolic constraints required to reproduce the results using in silico metabolic simulations. We found that the measured flux distribution can be reproduced by maximizing the ATP consumption in the flux balance analysis considering a limitation of metabolic heat dissipation (enthalpy change). Consistent with the simulation, OXPHOS inhibition induced metabolic redirection to aerobic glycolysis while maintaining the intracellular temperature. Furthermore, the dependency on aerobic glycolysis was partly alleviated upon culturing at low temperatures. Our data suggest that metabolic thermogenesis is an important factor in understanding aerobic glycolysis in cancer cells and that an advantage of aerobic glycolysis is the reduction in metabolic heat generation during ATP regeneration.

通过底物水平磷酸化再生三磷酸腺苷（ATP）是癌症代谢的一个普遍特征，即使在常氧条件下（有氧糖酵解）。然而，目前尚不清楚为什么癌细胞更喜欢低效的有氧糖酵解而不是高效的氧化磷酸化过程来再生ATP。为了研究有氧糖酵解的代谢原理，我们对12株培养的癌细胞进行了13c代谢通量分析，并探索了利用硅代谢模拟重现结果所需的代谢限制。我们发现，考虑到代谢热耗散（焓变）的限制，通量平衡分析中可以通过最大化ATP消耗来再现测量的通量分布。与模拟一致，OXPHOS抑制诱导代谢重定向到有氧糖酵解，同时保持细胞内温度。此外，低温培养部分减轻了对有氧糖酵解的依赖。我们的数据表明，代谢产热是理解癌细胞中有氧糖酵解的一个重要因素，有氧糖酵解的一个优势是在ATP再生过程中代谢产热的减少。

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引用次数: 0

Developing a Redox Imbalance Forces Drive (RIFD) strategy and its application in L-threonine production 氧化还原不平衡力驱动（RIFD）策略及其在l -苏氨酸生产中的应用。

IF 6.8 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

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

Xin Jin , Ruxin Hao , Hannuo Shen , Zhu Liu , Sumeng Wang , Qingsheng Qi , Quanfeng Liang

The design of synthetic driving forces for biosynthetic pathway is crucial for directing carbon flux toward the target product. Optimizing cellular redox status is one of the key strategies for constructing microbial cell factories. In this study, we attempt to create a novel redox imbalance force-driven (RIFD) strategy to direct carbon flow toward the target synthetic pathway. Initially, we increased the NADPH pool through a strategy of “open source and reduce expenditure” employing four approaches to achieve excessive NADPH levels and growth inhibition: (I) the expression of cofactor-converting enzymes, (II) the expression of heterologous cofactor-dependent enzymes, (III) the expression of enzymes involved in the NADPH synthesis pathway, and (IV) reduced NADPH wastage by knocking down non-essential genes that consume NADPH in vivo. Next, multiple automated genome engineering (MAGE) techniques were employed to evolve redox-imbalanced engineered strains and drive the metabolic flux to L-threonine production. Finally, we developed a NADPH and L-threonine dual-sensing biosensor, combined it with Fluorescence-Activated Cell Sorting (FACS), and a high-yield (0.65 g/g) L-threonine-producing strain with a titer of 117.65 g L⁻¹ was obtained. This research presents a general approach to increasing the production of cofactor-related products. Utilizing redox imbalance forces to drive metabolic flow toward the target product, it is possible to increase production while simultaneously restoring cell growth.

生物合成途径合成驱动力的设计是将碳通量导向目标产物的关键。优化细胞氧化还原状态是构建微生物细胞工厂的关键策略之一。在本研究中，我们试图创建一种新的氧化还原不平衡力驱动（RIFD）策略，将碳流导向目标合成途径。最初，我们通过“开源和减少支出”的策略增加了NADPH池，采用四种方法来实现过量的NADPH水平和生长抑制：(I)表达辅酶转换酶，（II）表达异源辅酶依赖酶，（III）表达参与NADPH合成途径的酶，（IV）通过敲除体内消耗NADPH的非必需基因来减少NADPH的浪费。接下来，采用多种自动化基因组工程（MAGE）技术来进化氧化还原不平衡的工程菌株，并驱动l -苏氨酸生产的代谢通量。最后，我们开发了一种NADPH和L-苏氨酸双传感生物传感器，并将其与荧光活化细胞分选（FACS）技术结合，获得了一株L-苏氨酸高产菌株（0.65 g/g），滴度为117.65 g/L。本研究提出了增加辅因子相关产品生产的一般方法。利用氧化还原不平衡力来驱动代谢流向目标产物，有可能在增加产量的同时恢复细胞生长。

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引用次数: 0

Genome mining of tailoring enzymes from biosynthetic gene clusters for synthetic biology: A case study with fungal methyltransferases. 从合成生物学的生物合成基因簇中挖掘裁剪酶的基因组：真菌甲基转移酶的案例研究。

IF 8.4 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

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

Liwen Zhang,Yang Liu,Kang Chen,Qun Yue,Chen Wang,Linan Xie,István Molnár,Yuquan Xu

Harnessing the potential of tailoring enzymes within fungal natural product (NP) biosynthetic gene clusters (BGCs) can significantly enhance NP diversity and production efficiency via artificially constructed microbial cell factories. To achieve this, an efficient genome mining method is crucial, especially since the functions of many putative enzymes in databases are unknown. As a test case, we aimed to identify methyltransferases (MTs) that modify a polyketide substrate without a known cognate MT. 16,748 putative MTs were annotated in 101,321 fungal BGCs and grouped into orthologous families. Three methods were explored to prioritize suitable enzymes. Among these, the machine learning method proved superior, with 11 out of 15 tested MTs successfully methylating the test substrate. This demonstrates the effectiveness of machine learning to mine tailoring enzymes that modify selected compounds, aiding synthetic biology in optimizing NP biosynthesis and facilitating the production of "unnatural products" for pharmaceutical or other bioindustrial applications.

利用真菌天然产物（NP）生物合成基因簇（BGCs）中剪裁酶的潜力，可以通过人工构建微生物细胞工厂显著提高NP多样性和生产效率。为了实现这一点，一种有效的基因组挖掘方法是至关重要的，特别是因为数据库中许多假定的酶的功能是未知的。作为一个测试案例，我们旨在鉴定甲基转移酶（MTs）修饰聚酮底物而没有已知的同源MT。在101,321个真菌BGCs中注释了16,748个假定的MTs，并将其分为同源家族。探索了三种优选合适酶的方法。其中，机器学习方法被证明是优越的，15个被测试的mt中有11个成功地甲基化了测试底物。这证明了机器学习在挖掘修饰选定化合物的剪裁酶方面的有效性，帮助合成生物学优化NP生物合成，并促进用于制药或其他生物工业应用的“非天然产品”的生产。

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引用次数: 0

Targeted metabolomics-guided rational refinement of cyanobacterial metabolism enables enhanced photosynthetic production of L-lysine. 有针对性的代谢组学指导蓝藻代谢的合理细化，使l -赖氨酸的光合作用生产增强。

IF 8.4 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

Pub Date : 2025-08-01 DOI: 10.1016/j.ymben.2025.07.015

Bo Wang,Piyoosh K Babele,Miles N Crockett,Joshua P Abraham,Sara Weidenbach,Brian F Pfleger,Jamey D Young

Cyanobacteria are capable of fixing CO2 using sunlight as the sole energy source and are promising microbial platforms for sustainable bioproduction of fuels, commodity chemicals, food and pharmaceuticals. L-lysine is an essential amino acid to humans and animals and is a precursor to synthesis of building blocks for nylon and polyesters. Its industrial production is currently solely based upon fermenting sugars by heterotrophic microorganisms such as Corynebacterium and Escherichia coli. Engineering cyanobacteria to effectively redirect photosynthetically fixed carbon towards lysine biosynthesis would provide a carbon-negative route of lysine production. In this study, to address the bottlenecks of lysine biosynthesis in the cyanobacterium Synechococcus sp. PCC 7002 (also called Picosynechococcus sp. PCC 7002), we combined targeted metabolomics and synthetic biology approaches to progressively identify and mitigate the major rate-limiting steps, ultimately increasing lysine productivity by about 77% compared to the parental strain. The best engineered lysine-producing Synechococcus strain was able to produce up to 2.92 mM lysine within 5 days, with flux partitioning towards lysine peaking at 23% of total fixed carbon under phototrophic conditions. The strategies demonstrated in this study can be used to guide rational metabolic engineering to identify and overcome pathway bottlenecks that limit flux to other renewable bioproducts in cyanobacteria.

蓝藻能够利用阳光作为唯一的能源来固定二氧化碳，是燃料、商品化学品、食品和药品可持续生物生产的有前途的微生物平台。赖氨酸是人类和动物必需的氨基酸，是合成尼龙和聚酯材料的前体。它的工业生产目前完全基于异养微生物如棒状杆菌和大肠杆菌发酵糖。工程蓝藻有效地将光合作用固定碳转向赖氨酸生物合成将提供赖氨酸生产的碳负途径。在本研究中，为了解决蓝藻聚球菌sp. PCC 7002（也称为皮聚球菌sp. PCC 7002）中赖氨酸生物合成的瓶颈，我们结合了靶向代谢组学和合成生物学方法，逐步识别和减轻主要的限速步骤，最终使赖氨酸的产量比亲本菌株提高约77%。最佳的工程产赖氨酸聚球菌菌株在5天内可产生2.92 mM赖氨酸，在光养条件下，赖氨酸的通量分配达到总固定碳的23%。本研究中展示的策略可用于指导合理的代谢工程，以识别和克服限制蓝藻中其他可再生生物产品通量的途径瓶颈。

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引用次数: 0

De novo biosynthesis of D-panthenol in engineered E. coli with rationally designed L-homoserine decarboxylase 用合理设计的l -丝氨酸脱羧酶在工程大肠杆菌中重新合成d -泛醇

IF 6.8 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

Pub Date : 2025-07-28 DOI: 10.1016/j.ymben.2025.07.013

Peng Zheng , Jie Ren , Jie Zheng , Feixia Liu , Xinhao Han , Bo Yu

D-panthenol is a compound of significant importance in the pharmaceutical, cosmetic, and nutraceutical sectors, attributed to its remarkable moisturizing, anti-inflammatory, and tissue repair properties. Traditional chemical synthesis encounters several challenges, including the generation of toxic by-products, low enantiomeric excess, and expensive purification processes. To date, complete biosynthesis of D-panthenol solely from glucose has seldom been documented. In this study, we have developed a new fermentative route to produce D-panthenol. The pathway incorporates previously unreported reaction of decarboxylating L-homoserine to 3-amino-1-propanol, which is achieved by rational design of a novel tyrosine decarboxylase mutant, informed by structural and mechanistic insights into enzymes acting on sterically similar substrates. The next enzyme facilitating the condensation of 3-amino-1-propanol with D-pantoate for D-panthenol formation was identified through a comprehensive screening of natural D-pantothenate synthetases. The artificial pathway was functionally expressed in a minimally engineered E. coli strain, resulting in the de novo production of D-panthenol from glucose. This research highlights a demonstration of an unnatural enzymatic synthesis process for D-panthenol. With further strain and process engineering, this new approach could be a promising way to produce D-panthenol biologically.

d -泛醇是一种在制药、化妆品和营养保健领域具有重要意义的化合物，因其具有显著的保湿、抗炎和组织修复特性。传统的化学合成遇到了一些挑战，包括产生有毒的副产物，低对映体过剩，和昂贵的净化过程。迄今为止，仅由葡萄糖完全生物合成d -泛醇的文献很少。在本研究中，我们开发了一种新的生产d -泛醇的发酵途径。该途径结合了以前未报道的l -高丝氨酸脱羧到3-氨基-1-丙醇的反应，这是通过合理设计一种新型酪氨酸脱羧酶突变体实现的，该突变体通过对作用于空间相似底物的酶的结构和机制的了解来实现的。下一个促进3-氨基-1-丙醇与d -泛酸酯缩合形成d -泛酸酯的酶是通过对天然d -泛酸酯合成酶的综合筛选确定的。人工途径在一种最低限度工程的大肠杆菌菌株中功能性表达，导致从葡萄糖中重新产生d -泛醇。这项研究强调了一个非自然的d -泛醇酶合成过程的演示。通过进一步的菌株和工艺工程，这种新方法可能是一种有前途的生物生产d -泛醇的方法。

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引用次数: 0

Sustainable production through spatial niche partitioning in engineered light-driven microbial community 工程光驱动微生物群落空间生态位分配的可持续生产

IF 6.8 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

Pub Date : 2025-07-28 DOI: 10.1016/j.ymben.2025.07.012

Hao Gao , Yifan Song , Yujia Jiang , Wankui Jiang , Feng Guo , Ziyi Yu , Minjiao Chen , Guodong Luan , Jee Loon Foo , Wenming Zhang , Matthew Wook Chang , Fengxue Xin , Min Jiang

Light-driven microbial communities consisting of phototrophs and heterotrophs represent an emerging frontier for biochemicals production from carbon dioxide (CO₂). However, the construction of stable and robust light-driven artificial microbial communities remains challenging because the dominant strain wins the competition for nutrient and leads to the instability of subpopulations. Inspired by natural ecosystems, one promising approach to assemble stable consortia is to construct spatial niches partitioning subpopulations—that is, physically separating different microbial members into distinct microenvironments to reduce competition and enable stable coexistence. Herein, a light-driven microbial community containing an autotrophic Synechococcus elongatus FL130 strain and a heterotrophic Meyerozyma guilliermondii strain was first constructed. Then, we developed spatially arranged core-shell microgels, enabling the precise control of subpopulations of different microbial members. Next, these microgels were integrated into macroscopic living material scaffold using extrusion bioprinting to advance bioprocessing applications, obtaining a well-coupled, robust and reusable light-driven microbial community. This resulted in a light-driven microbial communities with spatially compartmentalized distribution that can efficiently convert CO₂ into valuable chemical products of 2-phenylethanol and tyrosol, representing a pioneering approach for sustainable high-value biochemical production.

由光养生物和异养生物组成的光驱动微生物群落代表了二氧化碳（CO2）生物化学生产的新兴前沿。然而，由于优势菌株赢得营养竞争并导致亚群的不稳定，构建稳定而强健的光驱动人工微生物群落仍然具有挑战性。受自然生态系统的启发，构建空间生态位划分亚种群是构建稳定群落的一种很有希望的方法，即物理上将不同的微生物成员分离到不同的微环境中，以减少竞争，实现稳定的共存。本文首先构建了一个包含自养长聚球菌FL130菌株和异养吉列mondii Meyerozyma菌株的光驱动微生物群落。然后，我们开发了空间排列的核壳微凝胶，可以精确控制不同微生物成员的亚群。接下来，利用挤出生物打印技术将这些微凝胶整合到宏观生物材料支架中，以推进生物加工应用，获得一个耦合良好、健壮且可重复使用的光驱动微生物群落。这就形成了一个具有空间分区分布的光驱动微生物群落，可以有效地将二氧化碳转化为有价值的2-苯乙醇和酪醇的化学产物，代表了可持续高价值生化生产的开创性方法。

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引用次数: 0

Quantifying supply and demand in the pea aphid-Buchnera symbiosis reveals the metabolic Achilles’ heels of this interaction 定量的供应和需求在豌豆蚜虫- buchnera共生揭示了代谢的阿基里斯之踵，这种相互作用。

IF 6.8 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

Pub Date : 2025-07-26 DOI: 10.1016/j.ymben.2025.07.011

Léo Gerlin, Karen Gaget, Garance Lapetoule, Yohann Quivet, Patrice Baa-Puyoulet, Isabelle Rahioui, Mélanie Ribeiro Lopes, Pedro Da Silva, Federica Calevro, Hubert Charles

Many herbivorous insects feed on unbalanced diets and rely on bacterial endosymbionts to meet all their nutritional needs. This is the case for the pea aphid (Acyrthosiphon pisum), a plant pest whose remarkable growth and reproductive capacities cannot be sustained by its sole nutritional resource, the plant phloem sap, and which relies on a symbiotic relationship maintained over millions of years with the intracellular bacterium Buchnera aphidicola for the biosynthesis of amino acids and vitamins. Exploiting original experimental data and metabolic reconstructions, we have built a quantitative genome-scale metabolic model of B. aphidicola and used it to quantify amino acid exchanges between the bacterium and its host. We found metabolites that can rewire pathways, influencing the balance between selfish (growth-focused) and mutualist (amino acid synthesis) behavior. Among the products synthesized by Buchnera, phenylalanine, tyrosine and leucine are the main matter sinks and consume more than 60 % of imported glucose and serine. Finally, we compared the predicted bacterial supply to the aphid demand in amino acids. We found that the pea aphid may efficiently regulate its symbiont population density depending on its metabolic requirements, but that embryos are quantitatively not self-sustaining, with embryonic bacteria supply falling short of demand by 50 %. Overall, our study highlights candidate compounds and pathways to target for destabilizing this symbiosis or predicting its resilience to environmental or nutritional perturbations.

许多草食性昆虫以不平衡的饮食为食，依靠细菌内共生体来满足它们所有的营养需求。豌豆蚜虫（Acyrthosiphon pisum）就是这种情况，这种植物害虫的显著生长和繁殖能力不能靠其唯一的营养来源——植物韧皮部汁液来维持，它依赖于与细胞内细菌蚜虫（Buchnera aphidicola）维持了数百万年的共生关系来合成氨基酸和维生素。利用原始实验数据和代谢重建，我们建立了一个定量的蚜虫基因组尺度的代谢模型，并利用它来量化细菌与宿主之间的氨基酸交换。我们发现代谢物可以重新连接通路，影响自私（以生长为中心）和互惠（氨基酸合成）行为之间的平衡。在Buchnera合成的产品中，苯丙氨酸、酪氨酸和亮氨酸是主要的物质汇，消耗了60%以上的进口葡萄糖和丝氨酸。最后，我们比较了预测的细菌供应和蚜虫对氨基酸的需求。我们发现豌豆蚜虫可以根据其代谢需求有效地调节其共生体的种群密度，但胚胎在数量上不能自我维持，胚胎细菌供应不足需求的50%。总的来说，我们的研究突出了候选化合物和途径，以破坏这种共生关系或预测其对环境或营养扰动的恢复能力。

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引用次数: 0

Enhancing ε-poly-L-lysine production in Streptomyces albulus through L-lysine importer engineering 利用l -赖氨酸进口工程提高白球链霉菌的ε-聚l -赖氨酸产量。

IF 6.8 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Metabolic engineering

Pub Date : 2025-07-25 DOI: 10.1016/j.ymben.2025.07.010

Daojun Zhu , Jiawei Zhang , Shangyu Li, Liang Wang, Hongjian Zhang, Jianhua Zhang, Zhang, Xusheng Chen

ε-Poly-L-lysine (ε-PL) is a homopolymer of L-lysine residues produced by microorganisms, widely utilized in the food, pharmaceutical, and cosmetic industries. However, the development of efficient microbial cell factories (MCFs) for ε-PL production remains challenging. In this study, L-lysine importers were systematically screened, identified, and engineered to enhance ε-PL biosynthesis. First, an ε-PL-producing strain, Streptomyces albulus GS114, efficiently utilizing exogenous L-lysine, was selected. Bioinformatics analysis identified seven putative L-lysine importers in GS114, among which GL6157 was confirmed as the primary importer through molecular docking, transcriptional analysis, and genetic manipulation. Through combinatorial optimization of GL6157 expression coupled with overexpression of ε-poly-L-lysine synthase (pls), we engineered the GS114/pls-GL6157 strain, which achieved a ε-PL of 94.0 g/L in fed-batch fermentation. To our knowledge, this represents the highest reported yield to date. These findings demonstrate that transporter engineering is an effective strategy for enhancing ε-PL biosynthesis in industrial MCFs.

ε-聚l -赖氨酸（ε-PL）是一种由微生物产生的l -赖氨酸残基的均聚物，广泛应用于食品、制药和化妆品等行业。然而，开发高效的微生物细胞工厂（mcf）生产ε-PL仍然具有挑战性。在本研究中，l -赖氨酸导入物被系统筛选、鉴定和改造以促进ε-PL的生物合成。首先，选择了一株能高效利用外源l -赖氨酸的产ε- pl菌株——白链霉菌GS114。生物信息学分析在GS114中鉴定出7个假定的l -赖氨酸进口蛋白，通过分子对接、转录分析和基因操作确定GL6157为主要进口蛋白。通过对GL6157的表达和过表达ε-聚L-赖氨酸合成酶（pls）的组合优化，构建了菌株GS114/pls-GL6157，该菌株在补料分批发酵条件下的ε-PL为94.0 g/L。据我们所知，这是迄今为止报道的最高产量。这些发现表明，转运体工程是促进工业MCFs中ε-PL生物合成的有效策略。

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