Metabolic engineering最新文献_第4页

Systems-level analysis provides insights on methanol-based production of l-glutamate and its decarboxylation product γ-aminobutyric acid by Bacillus methanolicus 系统级分析提供了甲醇芽孢杆菌以甲醇为基础生产l-谷氨酸及其脱羧产物γ-氨基丁酸的见解。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-06-10 DOI: 10.1016/j.ymben.2025.06.002

Marta Irla , Ingemar Nærdal , David Virant , Trygve Brautaset , Tobias Busche , Dušan Goranovič , Carsten Haupka , Stéphanie Heux , Gregor Kosec , Christian Rückert-Reed , Volker F. Wendisch , Luciana F. Brito , Cláudia M. Vicente

{"title":"Systems-level analysis provides insights on methanol-based production of l-glutamate and its decarboxylation product γ-aminobutyric acid by Bacillus methanolicus","authors":"Marta Irla , Ingemar Nærdal , David Virant , Trygve Brautaset , Tobias Busche , Dušan Goranovič , Carsten Haupka , Stéphanie Heux , Gregor Kosec , Christian Rückert-Reed , Volker F. Wendisch , Luciana F. Brito , Cláudia M. Vicente","doi":"10.1016/j.ymben.2025.06.002","DOIUrl":"10.1016/j.ymben.2025.06.002","url":null,"abstract":"<div><div><em>Bacillus methanolicus</em> is the next workhorse in biotechnology using methanol, an alternative and economical one-carbon feedstock that can be obtained directly from carbon dioxide, as both carbon and energy source for the production of value-added chemicals. The wild-type strain <em>B. methanolicus</em> MGA3 naturally overproduces <span>l</span>-glutamate in methanol-based fed-batch fermentations. Here we generated a <em>B. methanolicus</em> strain exhibiting enhanced <span>l</span>-glutamate production capability through induced mutagenesis. To showcase the potential of this mutant strain, further metabolic engineering enabled the production of γ-aminobutyric acid (GABA) directly from <span>l</span>-glutamate during methanol fed-batch fermentations. Using a systems-level analysis, encompassing whole-genome sequencing, RNA sequencing, fluxome analysis and genome-scale metabolic modelling, we were able to elucidate the metabolic and regulatory adaptations that sustain the biosynthesis of these products. The metabolism of the mutant strain specifically evolved to prioritize energy conservation and efficient carbon utilization, culminating in increased product formation. These results and insights provide a foundation for further rational metabolic engineering and bioprocess optimization, enhancing the industrial viability of <em>B. methanolicus</em> for sustainable production of <span>l</span>-glutamate and its derivatives.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 389-404"},"PeriodicalIF":6.8,"publicationDate":"2025-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144285411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Systematic engineering of the sterol synthesis pathway for Saccharomyces cerevisiae promotes the efficient production of β-caryophyllene 对酿酒酵母甾醇合成途径进行系统工程改造，促进了β-石竹烯的高效生产。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-06-04 DOI: 10.1016/j.ymben.2025.06.004

Yapeng Zhang , Chenwen Liu , Wenqiang Li , Zhidong Ma , Bo Lv , Lei Qin , Chun Li

{"title":"Systematic engineering of the sterol synthesis pathway for Saccharomyces cerevisiae promotes the efficient production of β-caryophyllene","authors":"Yapeng Zhang , Chenwen Liu , Wenqiang Li , Zhidong Ma , Bo Lv , Lei Qin , Chun Li","doi":"10.1016/j.ymben.2025.06.004","DOIUrl":"10.1016/j.ymben.2025.06.004","url":null,"abstract":"<div><div>β-caryophyllene, a plant-derived sesquiterpene, serves as a food flavoring, anti-inflammatory agent, antioxidant, and high-energy fuel source. Extraction of β-caryophyllene from plants is a costly and inefficient process. Therefore, microbial cell factories have been employed for the production of β-caryophyllene. However, the limited yield is insufficient for its industrial application. In this study, we balanced the utilization of cellular resources for growth and production by systematically regulating the sterol synthesis pathway to maximize the synthesis of β-caryophyllene. In the competitive pathways concerning sterol and fatty acid synthesis, genes expression was suppressed by substituting the original promoters with a glucose-sensing promoter P<sub><em>HXT1</em></sub> and a sterol synthesis promoter P<sub><em>ERG7</em></sub>, respectively. This approach effectively increased the production of β-caryophyllene by 6.8 times, reaching 854.7 mg/L. Engineering glucose-sensing pathway altered the strength of P<sub><em>HXT1</em></sub>, resulting in an increase in β-caryophyllene production to 1.25 g/L. The cell growth and β-caryophyllene production were further boosted through diploid fusion, resulting in 21.4 g/L β-caryophyllene in fed-batch fermentation. This represents the highest reported production of β-caryophyllene to date. This study provides a valuable reference for the production of sesquiterpenes in microbial cell factories.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 347-355"},"PeriodicalIF":6.8,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144248710","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Regulation of proenzyme activation and metabolic engineering for protein-glutaminase production in Bacillus subtilis 枯草芽孢杆菌蛋白谷氨酰胺酶原活化调控及代谢工程研究。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-06-02 DOI: 10.1016/j.ymben.2025.06.001

Maofang Teng , Juan Zhang , Jingwen Zhou , Jianghua Li , Guocheng Du , Jian Chen , Guoqiang Zhang

{"title":"Regulation of proenzyme activation and metabolic engineering for protein-glutaminase production in Bacillus subtilis","authors":"Maofang Teng , Juan Zhang , Jingwen Zhou , Jianghua Li , Guocheng Du , Jian Chen , Guoqiang Zhang","doi":"10.1016/j.ymben.2025.06.001","DOIUrl":"10.1016/j.ymben.2025.06.001","url":null,"abstract":"<div><div>The protein-glutaminase (PG, EC 3.5.1.44) specifically targets glutamine residues in proteins and peptides, and has significant potential for enhancing the functional characteristics and processing efficiency of plant proteins. However, natural PG production faces challenges such as low enzymatic yield and difficult genetic manipulation. To address these challenges, a novel self-activating PG expression system was developed in <em>Bacillus subtilis</em>. First, pro-PG (PPG)-activated proteases were identified in <em>B. subtilis</em> by constructing a series of engineered strains. Second, the co-expression of PPG and PPG-activated protease in <em>B. subtilis</em> WB800 for mature PG (mPG) production was analyzed, and it was found that the supply and activation of PPG during fermentation was insufficient. Therefore, the gene expression components of PPG and protease, including the promoter and RBS, were further optimized. In addition, the key genes of the maltose metabolic pathway were knocked out, and the engineered strain W8ΔM2-AE-Pmal380 showed the highest capacity for PG production. Finally, a 53.0 U/mL mPG yield was achieved in a 5-L bioreactor within 64 h. This study establishes an efficient platform for industrial PG production and provides a reference for the expression and activation of other proenzymes.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 336-346"},"PeriodicalIF":6.8,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144225885","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Improving metabolic engineering design with enzyme-thermo optimization 用酶热优化改进代谢工程设计。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-06-01 DOI: 10.1016/j.ymben.2025.05.009

Wenqi Xu , Jingyi Cai , Wenjun Wu , Qianqian Yuan , Zhitao Mao , Hongwu Ma

{"title":"Improving metabolic engineering design with enzyme-thermo optimization","authors":"Wenqi Xu , Jingyi Cai , Wenjun Wu , Qianqian Yuan , Zhitao Mao , Hongwu Ma","doi":"10.1016/j.ymben.2025.05.009","DOIUrl":"10.1016/j.ymben.2025.05.009","url":null,"abstract":"<div><div>Metabolic target and strategy design play a critical role in enhancing the DBTL (Design-Build-Test-Learn) cycle in metabolic engineering. Classical stoichiometric algorithms such as OptForce<sup>Must</sup> and FSEOF narrow the experimental search space but fail to account for thermodynamic feasibility and enzyme-usage costs, leaving a space for their predictive performance. In this study, we introduce ET-OptME, a framework integrating two algorithms that systematically incorporate enzyme efficiency and thermodynamic feasibility constraints into genome-scale metabolic models. By mitigating thermodynamic bottlenecks and optimizing enzyme usage through a stepwise constraint-layering approach, ET-OptME delivers more physiologically realistic intervention strategies when compared with experimental records. Quantitative evaluation of five product targets in the <em>Corynebacterium glutamicum</em> model reveals that the algorithm showing at least 292 %, 161 % and 70 % increase in minimal precision and at least 106 %, 97 % and 47 % increase in accuracy when compared with stoichiometric methods, thermodynamic constrained methods, and enzyme constrained algorithms respectively.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 356-365"},"PeriodicalIF":6.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144216319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Efficient biosynthesis of gallic acid by a syntrophic Escherichia coli co-culture system 大肠杆菌共培养系统高效合成没食子酸的研究。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-05-16 DOI: 10.1016/j.ymben.2025.05.005

Shucai Wang , Zexi Kong , Xuecheng Xu , Jian Zhang , Liangcai Lin , Fengli Wu , Qinhong Wang

{"title":"Efficient biosynthesis of gallic acid by a syntrophic Escherichia coli co-culture system","authors":"Shucai Wang , Zexi Kong , Xuecheng Xu , Jian Zhang , Liangcai Lin , Fengli Wu , Qinhong Wang","doi":"10.1016/j.ymben.2025.05.005","DOIUrl":"10.1016/j.ymben.2025.05.005","url":null,"abstract":"<div><div>Gallic acid (GA), a natural phenolic acid antioxidant, has significant therapeutic and industrial applications. However, its traditional manufacturing approach, based on plant extraction, has been associated with risks of environmental pollution as well as a limited range of applications. Consequently, microbial-based production of GA, being more environmental-friendly, is viewed as a potential alternative. This study reports the efficient biosynthesis of GA from renewable glucose via a syntrophic <em>Escherichia coli</em> co-culture system. An effective GA biosynthesis pathway was first analyzed and determined. Then the rate-limiting step involving the hydroxylation of protocatechuic acid (PCA) to GA was removed by integrating multiple copies of the key gene <em>pobA</em><sup>T294A/Y385F</sup> into the chromosome of a PCA-overproducing strain. The resulting strain GA10 produced 41.88 g/L GA with a yield of 0.185 mol/mol, but up to 9.54 g/L of the intermediate PCA accumulated in the fermentation broth. To overcome this issue, a catalytic strain COT03 was constructed by coupling the metabolism of excess intracellular NADPH supply with the NADPH-consuming reaction catalyzed by PobA<sup>T294A/Y385F</sup>. This yielded a syntrophic <em>E</em>. <em>coli</em> co-culture system that consisted of a GA-overproducing strain (GA10) and a growth-coupled biocatalytic strain (COT03). Following optimization of the culture conditions, the co-culture system produced 57.66 g/L GA from glucose within 75 h, with a yield of 0.233 mol/mol and an average productivity of 0.769 g/L/h. This study lays the foundation for the potential industrial biomanufacturing of GA from glucose.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 313-321"},"PeriodicalIF":6.8,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144087525","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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 for enhanced lipid productivity in nutrient-rich conditions: A scalable approach to microbial lipid production 在营养丰富的条件下提高脂质产量的工程解脂耶氏菌：一种可扩展的微生物脂质生产方法。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-05-13 DOI: 10.1016/j.ymben.2025.05.004

Dongpil Lee , Hyemin Park , Jae-Eung Kim , Yeonsoo Kim , Joo Hyun Park , Hyesoo Lee , Byoung Hoon Yoon , Boyoung Han , Joon Young Jung , Seungwoo Cha , Peter Lee , Ji-Sook Hahn

{"title":"Engineering Yarrowia lipolytica for enhanced lipid productivity in nutrient-rich conditions: A scalable approach to microbial lipid production","authors":"Dongpil Lee , Hyemin Park , Jae-Eung Kim , Yeonsoo Kim , Joo Hyun Park , Hyesoo Lee , Byoung Hoon Yoon , Boyoung Han , Joon Young Jung , Seungwoo Cha , Peter Lee , Ji-Sook Hahn","doi":"10.1016/j.ymben.2025.05.004","DOIUrl":"10.1016/j.ymben.2025.05.004","url":null,"abstract":"<div><div>Climate change is reducing crop yields and increasing price volatility for commodities like cocoa and palm oil, thereby driving the need for sustainable alternatives such as microbial lipid production. The oleaginous yeast <em>Yarrowia lipolytica</em> is a promising platform for lipid synthesis. However, its lipid accumulation has traditionally relied on nitrogen limitation, posing challenges for achieving high yields under nutrient-rich conditions. In this study, we engineered <em>Y. lipolytica</em> to enhance lipid accumulation and productivity in nutrient-rich environments. Key modifications included deleting <em>MHY1</em> to prevent filamentous growth, overexpressing triacylglycerol (TAG) biosynthetic genes, disrupting fatty acid degradation, and redirecting phosphatidic acid flux toward TAG biosynthesis by reducing phospholipid production through <em>OPI3</em> deletion and <em>CDS1</em> mutation. Furthermore, deletion of <em>CEX1</em> to block citrate excretion significantly enhanced lipid accumulation. The resulting strain, CJ0415, achieved a lipid production of 54.6 g/L with a lipid content of 45.8 % and a record lipid productivity of 2.06 g/L/h under nutrient-rich conditions in a 5-L fermenter, representing a 2.6-fold increase compared to nitrogen-limited conditions. These findings underscore the potential of <em>Y. lipolytica</em> as a robust platform for scalable, industrial lipid production under nutrient-rich conditions.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 302-312"},"PeriodicalIF":6.8,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144065812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Enhancing the production of isopropanol with reduced CO2 emission via protein and metabolic engineering using Corynebacterium glutamicum 利用谷氨酸棒状杆菌的蛋白质和代谢工程提高异丙醇的生产和减少二氧化碳排放。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-05-13 DOI: 10.1016/j.ymben.2025.05.006

Wu-Young Jeong , Eunseo In , Myeong-Eun Lee , Young Jin Ko , Sung Ok Han

{"title":"Enhancing the production of isopropanol with reduced CO2 emission via protein and metabolic engineering using Corynebacterium glutamicum","authors":"Wu-Young Jeong , Eunseo In , Myeong-Eun Lee , Young Jin Ko , Sung Ok Han","doi":"10.1016/j.ymben.2025.05.006","DOIUrl":"10.1016/j.ymben.2025.05.006","url":null,"abstract":"<div><div>Isopropanol (IPA), a versatile chemical with applications in various fields of industries, yet its petroleum-based production raises environmental concerns. In this study, <em>Corynebacterium glutamicum</em> was engineered to enhance IPA production while mitigating CO<sub>2</sub> emissions. First, the rational design of secondary alcohol dehydrogenase (SADH) variants with shifted cofactor specificity from NADPH to NADH, resulting in an 11.11-fold increased NADH oxidation rate and 6.02-fold increased enzyme activity. Secondly, modified SADH was used in combination with the <em>Ncgl1676</em> promoter in order to separate the growth and production phases. This engineering resulted in a strain called CGIPA-4, which showed a 2.45-fold increase in IPA production. To address CO<sub>2</sub> emission, carbonic anhydrase from <em>Hydrogenovibrio marinus</em> (<em>HmCA</em>) and acetoacetyl-CoA synthase (<em>nphT7</em>) were overexpressed, constructing CGIPA-5 strain, enabling conversion of CO<sub>2</sub> into bicarbonate, which supported IPA biosynthesis and reduced emissions by up to 21 %. Finally, high cell density fed-batch fermentation using CGIPA-5 strain produced 148.6 ± 3.8 g L<sup>−1</sup> IPA, with CO<sub>2</sub> emission reduced by 30 % compared to CGIPA-4 strain. This work demonstrates a sustainable approach to petrochemical replacement through protein and metabolic engineering.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 267-275"},"PeriodicalIF":6.8,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144065814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Metabolic engineering of acetogenic bacteria using CO gas-sensing transcriptional ON/OFF modules 利用CO气敏转录ON/OFF模块的产丙酮菌代谢工程。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-05-10 DOI: 10.1016/j.ymben.2025.04.012

Sangrak Jin , Irisappan Ganesh , Jiyun Bae , Donghwi Lee , Seulgi Kang , Hyeonsik Lee , Jeong Wook Lee , Byung-Kwan Cho

{"title":"Metabolic engineering of acetogenic bacteria using CO gas-sensing transcriptional ON/OFF modules","authors":"Sangrak Jin , Irisappan Ganesh , Jiyun Bae , Donghwi Lee , Seulgi Kang , Hyeonsik Lee , Jeong Wook Lee , Byung-Kwan Cho","doi":"10.1016/j.ymben.2025.04.012","DOIUrl":"10.1016/j.ymben.2025.04.012","url":null,"abstract":"<div><div>Dynamic sensing of gas substrates like toxic carbon monoxide (CO) in living microbial cells is often limited due to the lack of suitable biosensors. Here, we integrated the CO-binding transcription activators, CooA and RcoM1, with an O<sub>2</sub>-independent fluorescent reporter system, Halo-tag, to develop CO-sensing modules (ON/OFF) capable of detecting CO concentrations in the strictly anaerobic acetogenic bacterium <em>Eubacterium limosum</em>. Furthermore, we employed CooA as the CO-sensing ON module to activate the target genes for 2,3-butanediol (2,3-BDO) biosynthesis, achieving a 1.7-fold increase in 2,3-BDO yield. These results indicate that the CO-ON module effectively redirects carbon flux toward target product biosynthesis pathway in acetogens. However, during CO gas with glucose mixotrophic fermentation, lactate emerged as the predominant product. To enhance target pathway flux using the CO-ON module, we deleted the lactate pathway in <em>E. limosum</em> using CRISPR/Cas9. The resulting engineered strain showed an 18.5 % increase in carbon utilization for 2,3-BDO production under CO sensing culture conditions. This optimized platform strain subsequently produced approximately 52 g/L of 2,3-BDO during two stage CO-glucose mixotrophic fermentation. Our results provide orthogonal CO-sensing transcriptional regulatory modules for engineering metabolic pathways that efficiently convert CO into value-added biochemicals using acetogenic biocatalysts.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 290-301"},"PeriodicalIF":6.8,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143945092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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 oleaginous yeast Rhodotorula toruloides for production of alkanes and alkenes 用于生产烷烃和烯烃的工程产油酵母。

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-05-07 DOI: 10.1016/j.ymben.2025.05.003

Yi Yu , Quan Yuan , Jing Dai , Huimin Zhao , Shuobo Shi

{"title":"Engineering oleaginous yeast Rhodotorula toruloides for production of alkanes and alkenes","authors":"Yi Yu , Quan Yuan , Jing Dai , Huimin Zhao , Shuobo Shi","doi":"10.1016/j.ymben.2025.05.003","DOIUrl":"10.1016/j.ymben.2025.05.003","url":null,"abstract":"<div><div>Due to limited reserves and excessive carbon emission of fossil fuels, there has been an increasing interest in developing advanced biofuels with high energy density such as alkanes and alkenes. Here we report the design and construction of three heterologous biosynthetic pathways of alkanes and alkenes in oleaginous yeast <em>Rhodotorula toruloides</em>, including the AAR/ADO, UndA/UndB and FAP pathways. The performance of various enzymes from different organisms was evaluated within <em>R. toruloides</em> for each pathway. Various metabolic engineering strategies were used to enhance the production of alkanes and alkenes across all three pathways, including enzyme screening, byproduct elimination, and precursor supply enhancement. Notably, the FAP pathway demonstrated significantly superior performance compared to the AAR/ADO and UndA/UndB pathway. As a result, 1.73 g/L alkanes and alkenes were produced from glucose, and 0.94 g/L alkanes and alkenes were produced from lignocellulosic hydrolysates, representing the highest alkanes and alkenes titers reported in yeast. This work establishes <em>R. toruloides</em> as a promising host for hydrocarbons production from glucose and CO<sub>2</sub>-neutral feedstocks and paves the way for further strain and process optimization towards industrial production of alkanes and alkenes.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 242-253"},"PeriodicalIF":6.8,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932519","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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

Photosynthetic sorbitol production in Synechococcus sp. PCC 7002 is enhanced by addressing phosphatase promiscuity, nutrient availability and Calvin cycle bottlenecks 聚球菌PCC 7002通过解决磷酸酶混杂性、养分有效性和卡尔文循环瓶颈，提高了光合作用山梨醇的产量

IF 6.8 1区生物学

Metabolic engineering Pub Date : 2025-05-02 DOI: 10.1016/j.ymben.2025.04.008

Cody Kamoku , Pranav Bhavaraju , Collin Travis , Luis Taquillo , David R. Nielsen

{"title":"Photosynthetic sorbitol production in Synechococcus sp. PCC 7002 is enhanced by addressing phosphatase promiscuity, nutrient availability and Calvin cycle bottlenecks","authors":"Cody Kamoku , Pranav Bhavaraju , Collin Travis , Luis Taquillo , David R. Nielsen","doi":"10.1016/j.ymben.2025.04.008","DOIUrl":"10.1016/j.ymben.2025.04.008","url":null,"abstract":"<div><div>Cyanobacteria represent promising biocatalysts for producing carbohydrates, including sorbitol, a naturally-occurring, fermentable sugar alcohol with conventional uses as a sweetener, pharmaceutical additive, and biodegradable plasticizer. Previously, <em>Synechocystis</em> sp. PCC 6803 was engineered to produce sorbitol, reaching a final titer of 2.3 g/L after 18 days. To improve upon this performance, sorbitol production was herein engineered in the faster growing strain <em>Synechococcus</em> sp. PCC 7002. Upon introducing the sorbitol biosynthetic pathway, up to 500 mg/L sorbitol was initially produced after seven days. However, due to the initial use of two highly promiscuous sugar phosphatase variants, this also resulted in the unwanted co-production of ribose and growth inhibition due to depletion of ribose-5-phosphate from the Calvin cycle. This off-target effect was ultimately mitigated via the discovery that mannitol-1-phosphate phosphatase from <em>Eimeria tenella</em> also dephosphorylates sorbitol-6-phosphate to sorbitol with greater specificity, leading to improved growth and sorbitol production. Next, two bottleneck enzymes in the Calvin cycle, namely fructose-bisphosphate aldolase (FBA) and bifunctional fructose-1,6-bisphosphatase/sedoheptulose-1,7-bisphosphatase (BiBPase), were overexpressed both individually and in combination, resulting in sorbitol production up to 1.3 g/L. Finally, upon optimizing the culture media to address nutrient limitation, the final strain produced up to 3.6 g/L sorbitol in nine days, respectively representing 1.5- and 3-fold increases in titer and productivity relative to previously-engineered <em>Synechocystis</em> sp. PCC 6803.</div></div>","PeriodicalId":18483,"journal":{"name":"Metabolic engineering","volume":"91 ","pages":"Pages 181-191"},"PeriodicalIF":6.8,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143911542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"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