Synthetic and Systems Biotechnology最新文献_第2页

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

IF 4.4 2区生物学

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

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

{"title":"Systems metabolic engineering of dual pathways with stage-specific activation for high-level 5-aminolevulinic acid production in Escherichia coli","authors":"Yanghao Liu , Changgeng Li , Yu Miao , Zhichao Chen , Tang'en Shi , Ling Ma , Siyu Zhao , Zichen Yu , Lanxiao Li , Qingyang Xu","doi":"10.1016/j.synbio.2025.09.007","DOIUrl":"10.1016/j.synbio.2025.09.007","url":null,"abstract":"<div><div>5-Aminolevulinic acid (5-ALA) is an important non-proteinogenic amino acid with wide applications in agriculture and medicine. To achieve high-level microbial production, this study established a dual-pathway reconstruction strategy in <em>Escherichia coli</em> by integrating the endogenous C5 pathway with an inducible exogenous C4 pathway. Multi-copy overexpression of <em>gltX</em>, <em>hemA</em>, and <em>hemL</em>, combined with enhanced glutamate supply and the introduction of a non-oxidative glycolysis pathway, effectively increased the C5 pathway flux and carbon efficiency, while product toxicity was significantly alleviated by strengthening efflux mechanisms and oxidative stress tolerance. A quorum sensing–based regulatory system was employed to dynamically regulate <em>hemB</em> expression, thereby balancing cell growth and 5-ALA biosynthesis, and a controlled glycine feeding strategy specifically activated the C4 pathway during the later fermentation stage, forming a stage-specific dynamic activation mechanism. Promoter regulation of <em>sucC/sucD</em> expression and enhanced endogenous PLP biosynthesis further stabilized the C4 flux. Fed-batch fermentation in a 5 L bioreactor resulted in a final 5-ALA titer of 37.34 g/L, demonstrating the industrial potential of this systems metabolic engineering strategy combining dual pathways and stage-specific activation control.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 141-151"},"PeriodicalIF":4.4,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145096567","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Engineering riboflavin-overproducing Bacillus subtilis via pathway gene overexpression 通过途径基因过表达工程制造核黄素过量的枯草芽孢杆菌

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2025-09-05 DOI: 10.1016/j.synbio.2025.09.005

Sijia Wang , Qiyao Zhu , Chuan Liu , Huina Dong , Miaomiao Xia , Zhaoxia Jin , Dawei Zhang

{"title":"Engineering riboflavin-overproducing Bacillus subtilis via pathway gene overexpression","authors":"Sijia Wang , Qiyao Zhu , Chuan Liu , Huina Dong , Miaomiao Xia , Zhaoxia Jin , Dawei Zhang","doi":"10.1016/j.synbio.2025.09.005","DOIUrl":"10.1016/j.synbio.2025.09.005","url":null,"abstract":"<div><div>Overexpression of genes at key metabolic nodes frequently enhances the yield of target products. However, this often leads to widespread issues with growth defect and plasmid instability. This study utilizes the riboflavin over-producer <em>Bacillus subtilis</em> U3 as a model to systematically investigate the impact of overexpressing riboflavin operon genes on strain growth and plasmid structural instability. Overexpression of genes in rib operon enhances target product yields by 13.2 % in U3, but the engineered strains suffer growth defects and plasmid instability. Frameshift mutations in the <em>ribD</em> gene were found to significantly reduce the loss of operon gene fragments by 16.7 %. Furthermore, this study investigates the application of the Respiration Activity Monitoring System (RAMOS) in evaluating the growth of metabolically engineered strains. RAMOS serves as a medium optimization tool and pre-fermentation screening platform. Specifically, guanine supplementation increased biomass by 11.1 % in <em>zwf</em>-overexpressing strains, whereas histidine, uracil, and tryptophan supplementation could improve the biomass of <em>purF</em>-overexpressing strain by 71.1 %. This approach provides a methodological framework for resolving growth defects in metabolic engineering.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 91-101"},"PeriodicalIF":4.4,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145061146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

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

IF 4.4 2区生物学

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

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

{"title":"Identification of a novel β-1,4-galactosyltransferase from Haemophilus parainfluenzae for efficient biosynthesis of lacto-N-neotetraose in Escherichia coli","authors":"Dan Liu , Chenhui Ying , Yingqi Ping , Shanquan Liang , Qiaojuan Yan , Zhengqiang Jiang","doi":"10.1016/j.synbio.2025.09.006","DOIUrl":"10.1016/j.synbio.2025.09.006","url":null,"abstract":"<div><div>As an important member of human milk oligosaccharides (HMOs), lacto-<em>N</em>-neotetraose (LNnT) has been recognized as a promising HMO with many physiological activities for infants. β-1,4-Galactosyltransferases play a vital role in the final step of LNnT biosynthesis in microbial cell factory. Herein, we identified a novel β-1,4-galactosyltransferase (Hpa-GalT) from <em>Haemophilus parainfluenzae</em> for the efficient biosynthesis of LNnT in <em>E. coli</em> BL21 star(DE3). After overexpression of the key genes in the synthetic pathway, the titer of LNnT reached 1.86 g/L in shake flask fermentation. Moreover, a variant A186P/E146Q of Hpa-GalT was obtained by molecular modification, resulting in an increase of the LNnT titer to 2.15 g/L. Subsequently, a series of genes related to competitive pathways were deleted, including <em>lacZ</em>, <em>ugd</em>, <em>wcaJ</em>, and <em>wecB</em>, the engineered strain produced a LNnT titer of 2.23 g/L. After optimization of LNnT transport along with LNT II utilization, the engineered strain AH31 produced 2.93 g/L of LNnT in shake flask fermentation. Furthermore, it produced a maximum titer of 32.6 g/L of total LNnT and 27.1 g/L of extracellular LNnT in a 5-L fermentor with the productivity for 0.52 g/L/h. Overall, this study provides a novel β-1,4-galactosyltransferase and lays a foundation for the efficient biosynthesis of LNnT.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 102-109"},"PeriodicalIF":4.4,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145061143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Engineering Yarrowia lipolytica as a yeast cell factory for the de novo production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) 利用酵母细胞工厂改造脂化耶氏菌，重新生产聚（3-羟基丁酸-co-4-羟基丁酸）

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2025-09-04 DOI: 10.1016/j.synbio.2025.09.002

Jinming Liu, Jia Yan, Zhiyong Cui, Qingsheng Qi

{"title":"Engineering Yarrowia lipolytica as a yeast cell factory for the de novo production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate)","authors":"Jinming Liu, Jia Yan, Zhiyong Cui, Qingsheng Qi","doi":"10.1016/j.synbio.2025.09.002","DOIUrl":"10.1016/j.synbio.2025.09.002","url":null,"abstract":"<div><div>The oleaginous yeast <em>Yarrowia lipolytica</em> presents a promising chassis for the production of biodegradable plastic polyhydroxyalkanoates (PHAs) due to its advantages in precursor supply capabilities and cost-effective substrate utilization. However, <em>Y. lipolytica</em> has primarily been engineered for the synthesis of homopolymeric PHAs, such as poly (3-hydroxybutyrate) (P3HB), which have limited material properties and application potential. In this study, the succinate dehydrogenase-deficient <em>Y. lipolytica</em> strains were utilized as hosts for the de novo biosynthesis of copolymer poly (3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB). By employing a strategy of subcellular partitioning, poly (4-hydroxybutyrate) (P4HB) was synthesized in the cytoplasm using 4-hydrobutyrate (4HB) derived from mitochondrial sources as a monomer. We then introduced the biosynthetic pathway for 3-hydroxybutyryl-CoA to facilitate P34HB accumulation. Importantly, the ratio of 4HB monomer in P34HB could be adjusted from 9.17 mol% to 45.26 mol% by varying medium components and carbon sources. Under fed-batch fermentation conditions in a 5 L bioreactor, the engineered strain achieved a P34HB titer of 18.61 g/L representing 19.18 % of cell dry weight (CDW). This study not only demonstrates the potential of <em>Y. lipolytica</em> for P34HB synthesis, but also expands the monomer pool available for PHAs in yeast cell factories.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 152-160"},"PeriodicalIF":4.4,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145096566","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

The coupling effect of Penicillium baileys W2 in the Aspergillus flavus inhibition and peanut growth promotion 贝利青霉W2抑制黄曲霉与促进花生生长的耦合效应

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2025-09-04 DOI: 10.1016/j.synbio.2025.09.003

Qianqian Wang , Shihua Shan , Quanxi Sun , Xiaobo Zhao , Cuiling Yuan , Yifei Mou , Juan Wang , Caixia Yan , Qi Wang , Qingchen Rui , Chunjuan Li

{"title":"The coupling effect of Penicillium baileys W2 in the Aspergillus flavus inhibition and peanut growth promotion","authors":"Qianqian Wang , Shihua Shan , Quanxi Sun , Xiaobo Zhao , Cuiling Yuan , Yifei Mou , Juan Wang , Caixia Yan , Qi Wang , Qingchen Rui , Chunjuan Li","doi":"10.1016/j.synbio.2025.09.003","DOIUrl":"10.1016/j.synbio.2025.09.003","url":null,"abstract":"<div><div><em>Aspergillus flavus</em> is a significant plant pathogen, and peanut crops are particularly vulnerable to aflatoxin contamination. This vulnerability underscores the need for more effective control methods. In this study, the strain <em>Penicillium baileys</em> W2 was isolated from the rhizosphere soils of healthy peanut seedlings. The fermentation extract exhibited concentration-dependent inhibition of pathogenic <em>A. flavus</em> growth, with a minimum inhibitory concentration (MIC) of 55 % and a minimum fungicidal concentration (MFC) of 60 %. Physiological data and transcriptome analysis demonstrated that the W2 fermentation supernatant inhibited <em>A. flavus</em> growth by disrupting membrane permeability. Metabolomics analysis identified active compounds, including propylparaben, taxifolin, and phloretin, which exhibited significant antagonistic effects against <em>A. flavus</em>. Additionally, we evaluated the impact of the W2 fermentation broth on peanut growth promotion and on rhizosphere microbial community structure using metagenomic sequencing. The reduction of harmful soil microorganisms contributed to the maintenance of soil health, whereas the increased abundance of beneficial microorganisms enhanced peanut seedling growth by facilitating soil nutrient cycling. These findings indicate that the development and application of <em>P. baileys</em> strain W2 or its fermentation extract aligns with sustainable agricultural principles and offers a promising biological control approach.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 127-140"},"PeriodicalIF":4.4,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145096568","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 vitamin B6 production in engineered Escherichia coli via restricted mix-carbon feeding and pressure-controlled fermentation 通过限制混合碳饲养和压力控制发酵提高工程大肠杆菌维生素B6的产量

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2025-09-04 DOI: 10.1016/j.synbio.2025.09.004

Long Chen , Miao-Miao Xia , Hui-Na Dong , Hao-Ran Ma , Xu-Yang Huang , Gong-Jin Shen , Zhao-Xia Jin , Lin-Xia Liu , Da-Wei Zhang

{"title":"Enhanced vitamin B6 production in engineered Escherichia coli via restricted mix-carbon feeding and pressure-controlled fermentation","authors":"Long Chen , Miao-Miao Xia , Hui-Na Dong , Hao-Ran Ma , Xu-Yang Huang , Gong-Jin Shen , Zhao-Xia Jin , Lin-Xia Liu , Da-Wei Zhang","doi":"10.1016/j.synbio.2025.09.004","DOIUrl":"10.1016/j.synbio.2025.09.004","url":null,"abstract":"<div><div>Pyridoxine (PN), a major commercial form of vitamin B<sub>6</sub>, is mainly produced via chemical synthesis, raising environmental concerns. Microbial fermentation offers a greener alternative, but low biomass and titer limit industrial application. In this study, we developed an integrated strategy combining DO-stat restricted mix-carbon feeding with two-stage pressure-controlled fermentation conditions, and medium optimization to enhance cell growth and PN production in <em>Escherichia coli</em>. DO-stat feeding enabled dynamic regulation of oxygen and carbon sources, while a two-stage pressure-controlled (0.1 bar for 0–24 h, 0.2 bar thereafter) improved both growth and PN biosynthesis. Meanwhile, a novel medium (CRS-67), optimized via Taguchi design, increased shake flask PN titer by 275 mg/L, 75.5 % over CS medium. Under systematic fermentation optimization in a 5-L bioreactor, OD<sub>600</sub> reached 142.8 and PN titer reached 3.33 g/L—the highest reported level to date. This study presents a robust and scalable process for microbial PN production and offers insights for industrial biomanufacturing of vitamin B<sub>6</sub>.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 76-84"},"PeriodicalIF":4.4,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145057208","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

Construction and analysis of a cell factory for terpenoid biosynthesis in Pichia pastoris via metabolic engineering and metabolomics 基于代谢工程和代谢组学的毕赤酵母萜类生物合成细胞工厂的构建与分析

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2025-09-03 DOI: 10.1016/j.synbio.2025.08.015

Chenfan Sun , Cuifang Ye , Xiaoqian Li , Jiabin Xu , Huiru Yu , Jucan Gao , Chengran Guan , Jintao Cheng

{"title":"Construction and analysis of a cell factory for terpenoid biosynthesis in Pichia pastoris via metabolic engineering and metabolomics","authors":"Chenfan Sun , Cuifang Ye , Xiaoqian Li , Jiabin Xu , Huiru Yu , Jucan Gao , Chengran Guan , Jintao Cheng","doi":"10.1016/j.synbio.2025.08.015","DOIUrl":"10.1016/j.synbio.2025.08.015","url":null,"abstract":"<div><div>Terpenoids are widely distributed in nature and have various applications in health products, pharmaceuticals, and fragrances. Despite the tremendous potential of terpenoids, traditional production methods such as plant extraction and chemical synthesis face challenges in meeting current market demands. With the developments in synthetic biology and metabolic engineering, it has become feasible to construct efficient microbial cell factories for large-scale production for terpenoids. In this work, using the yeast <em>Pichia pastoris</em> as the host cell, a \"plug-and-play\" cell factory for universal terpenoid production was constructed by enhancing the expression of the MVA pathway for common precursor synthesis and reducing branch pathway diversion strategies. We have successfully and efficiently synthesized β-elemene, β-farnesene, (+)-valencene, (−)-α-bisabolol by this cell factory. Furthermore, by analyzing metabolites in different engineered strains in terms of system biology, it was discovered that an increase in key protein copy numbers enhanced the synthesis of arginine and other metabolic pathways. The robustness of the strains and the tightly regulated metabolic network constrain rational metabolic engineering transformations. These data provide important clues for the modification and optimization of production strains.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 68-75"},"PeriodicalIF":4.4,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145046582","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

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

Synthetic and Systems Biotechnology Pub Date : 2025-09-02 DOI: 10.1016/j.synbio.2025.08.016

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

{"title":"A near-infrared ratio fluorescent probe achieves mitochondrial hydrogen polysulfide imaging for monitoring ferroptosis in arthritis","authors":"Ting Cao , Ziwen Xiao , Wenhua Dong , Hong Ma , Deyan Gong , Zhefeng Fan","doi":"10.1016/j.synbio.2025.08.016","DOIUrl":"10.1016/j.synbio.2025.08.016","url":null,"abstract":"<div><div>Reactive sulfur (RSS) is a type of sulfur-containing molecule widely present in biological systems. Hydrogen polysulfide (H<sub>2</sub>S<sub>n</sub>, 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 H<sub>2</sub>S<sub>n</sub> 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 H<sub>2</sub>S<sub>n</sub>. The experimental results show that the probe (<strong>Cy-S<sub>4</sub></strong>) 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 H<sub>2</sub>S<sub>n</sub> in ferroptosis process and fluorescence imaging of H<sub>2</sub>S<sub>n</sub> in inflammatory cell mitochondria. Pathological section data confirmed that the probe has good <em>in vivo</em> imaging ability, and more importantly, <em>in vivo</em> arthritis imaging experiments showed that the expression of H<sub>2</sub>S<sub>n</sub> 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.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 59-67"},"PeriodicalIF":4.4,"publicationDate":"2025-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145046581","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 multiplex metabolic engineering with adaptive evolution strategies for high-level succinic acid production in Yarrowia lipolytica 结合多元代谢工程和适应性进化策略的多脂耶氏菌高产琥珀酸的研究

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2025-08-30 DOI: 10.1016/j.synbio.2025.08.011

Tao Sun , Mei-Li Sun , Lu Lin , Jian Gao , Rodrigo Ledesma-Amaro , Kaifeng Wang , Xiao-Jun Ji

{"title":"Combining multiplex metabolic engineering with adaptive evolution strategies for high-level succinic acid production in Yarrowia lipolytica","authors":"Tao Sun , Mei-Li Sun , Lu Lin , Jian Gao , Rodrigo Ledesma-Amaro , Kaifeng Wang , Xiao-Jun Ji","doi":"10.1016/j.synbio.2025.08.011","DOIUrl":"10.1016/j.synbio.2025.08.011","url":null,"abstract":"<div><div>Succinic acid, an essential platform chemical with extensive utility in biodegradable materials, pharmaceuticals, and the food industry, faces challenges of high energy consumption and environmental pollution in traditional chemical synthesis. Here, we employed multiplex metabolic engineering and adaptive laboratory evolution to enhance succinic acid biosynthesis in <em>Y</em><em>arrowia lipolytica</em>. By attenuating succinate dehydrogenase (Sdh) activity, mitigating by-product accumulation, and enhancing the succinate synthesis pathway, engineered strains showed efficient succinic acid production from glycerol. The titer reached 130.99 g/L under unregulated pH conditions, translating to a yield of 0.35 g/g and a productivity of 0.70 g/(L·h). Subsequently, transporter engineering and adaptive evolution strategies were applied to enhance glucose utilization for succinic acid synthesis, yielding an evolved strain that eliminated the growth lag phase and produced 106.68 g/L succinic acid from glucose, which translated to a yield of 0.32 g/g and a productivity of 0.64 g/(L·h). Additionally, transcriptomic analysis and inverse metabolic engineering revealed that 4-hydroxyphenylpyruvate dioxygenase (4-Hppd) in the tyrosine degradation pathway partially restored the growth of Sdh-deficient strains on glucose, offering new insights for subsequent succinic acid biomanufacturing using <em>Y</em>. <em>lipolytica</em>.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 48-58"},"PeriodicalIF":4.4,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145027104","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

Development of a novel transformation system using zhongshengmycin in Aspergillus oryzae and Aspergillus niger 中生霉素在米曲霉和黑曲霉中转化体系的建立

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.synbio.2025.08.014

Futi Bi , Qichen Huang , Baoxiang Pan , Bin Wang , Li Pan

{"title":"Development of a novel transformation system using zhongshengmycin in Aspergillus oryzae and Aspergillus niger","authors":"Futi Bi , Qichen Huang , Baoxiang Pan , Bin Wang , Li Pan","doi":"10.1016/j.synbio.2025.08.014","DOIUrl":"10.1016/j.synbio.2025.08.014","url":null,"abstract":"<div><div>Filamentous fungi such as <em>Aspergillus oryzae</em> and <em>Aspergillus niger</em> serve as valuable microbial cell factories with diverse applications. However, genetic manipulation in these fungi is often hindered by the lack of efficient resistance-based transformation systems. To address this challenge, we developed a novel transformation system for both <em>A. oryzae</em> and <em>A. niger</em> based on zhongshengmycin, a cost-effective streptothricin-class antibiotic, used in combination with the resistance gene encoding nourseothricin acetyltransferase (NAT). Our tests revealed that sensitization with SDS and EDTA-Na<sub>2</sub> disrupts cell membrane integrity, thereby enhancing the susceptibility of <em>Aspergillus</em> to zhongshengmycin. Through an attB/attP-mediated integration system, we achieved heterologous expression of β-glucuronidase in <em>A. oryzae</em> with a selection efficiency of 85.71 % (6/7). Additionally, recombination of <em>megfp-sed5</em> in <em>A. niger</em> achieved a transformation efficiency of 83.33 % (10/12), comparable to that of the PyrG system, thus establishing a versatile platform for fungal biotechnology. This system enables precise genomic integration for metabolic rewiring, offering new strategies for the synthetic biology-driven improvement of industrial filamentous fungi.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"11 ","pages":"Pages 205-215"},"PeriodicalIF":4.4,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145220361","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