Biochemical Engineering Journal最新文献_第7页

Development of a high-efficiency constitutive expression system in Salinivibrio strain for next-generation extremophile engineering 新一代嗜极微生物工程中盐弧菌高效本构表达系统的建立

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-10 DOI: 10.1016/j.bej.2025.109863

Zheng-Jun Li, Nan Pu, Bin Wei, Hao Liang

{"title":"Development of a high-efficiency constitutive expression system in Salinivibrio strain for next-generation extremophile engineering","authors":"Zheng-Jun Li, Nan Pu, Bin Wei, Hao Liang","doi":"10.1016/j.bej.2025.109863","DOIUrl":"10.1016/j.bej.2025.109863","url":null,"abstract":"<div><div>The halophilic bacterium <em>Salinivibrio</em> sp. TGB10 represents an emerging microbial chassis for industrial biotechnology, yet its genetic toolkit remains underdeveloped due to insufficient characterization of regulatory elements. Here, we identified and characterized a novel endogenous promoter (p1) through transcriptomic analysis and functional validation. The 185-bp promoter region, located upstream of a gene encoding a hypothetical protein containing leucine-zipper lipoprotein domain, was found to contain all essential regulatory elements including a functional −10 element, extended −10 motif, and −35 element. Through progressive truncation and ribosome binding site engineering, we precisely mapped these elements and revealed their compensatory interactions in transcription initiation. Next, we developed a series of synthetic promoters by randomizing the spacer region between −35 and −10 elements, achieving expression levels spanning four orders of magnitude in <em>Salinivibrio</em> and three orders in <em>Escherichia coli</em>. The practical utility of p1 promoter was demonstrated through efficient production of industrially relevant β-glucosidase, with optimized fermentation conditions yielding 76.83 U/mL activity. This study provides a critical genetic toolkit for metabolic engineering and synthetic biology applications in <em>Salinivibrio</em>.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"223 ","pages":"Article 109863"},"PeriodicalIF":3.7,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144614317","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Detection of potential biomarkers for esophageal cancer based on fluorescent biosensor 基于荧光生物传感器的食管癌潜在生物标志物检测

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-10 DOI: 10.1016/j.bej.2025.109860

Liang Zhou , Tiantian Li , Shuaibing Yu , Pengnan Yao , Jinming Kong , Xueji Zhang

{"title":"Detection of potential biomarkers for esophageal cancer based on fluorescent biosensor","authors":"Liang Zhou , Tiantian Li , Shuaibing Yu , Pengnan Yao , Jinming Kong , Xueji Zhang","doi":"10.1016/j.bej.2025.109860","DOIUrl":"10.1016/j.bej.2025.109860","url":null,"abstract":"<div><div>Fluorescent biosensors have garnered significant interest due to their high sensitivity, efficiency, and rapid detection capabilities, making them an attractive alternative for various diagnostic applications. To address these needs, we developed a fluorescence-based biosensor using magnetic beads for the rapid and precise detection of miRNA-320b, a potential biomarker for esophageal cancer. The process begins by immobilizing DNA onto the surface of magnetic beads. Upon the introduction of miRNA-320b, a stable DNA-RNA duplex forms through complementary base pairing. In this biosensor design, 3-aminophenylboric acid serves as a \"crosslinker\" to connect carboxyfluorescein to miRNA-320b. The design relies on the cis-diol structure at the 3′ end of miRNA-320b, which forms a covalent bond with phenylboric acid via borate esterification, enabling the specific differentiation of miRNA-320b from DNA. The use of magnetic beads facilitates rapid separation and purification, minimizing interference and enhancing the detection of the esophageal cancer biomarker miRNA-320b. Following optimization, the biosensor displayed a linear range from 10⁻¹ ⁴ to 10⁻⁹ M with a minimum detection limit of 1.42 fM. This innovative fluorescent biosensor offers a novel, efficient, and straightforward approach for the early screening of esophageal cancer.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"223 ","pages":"Article 109860"},"PeriodicalIF":3.7,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144614320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Rational designing of human senescence marker protein 30 for enhanced hydrolysis of V-type nerve agent analog 人衰老标志蛋白30的合理设计促进v型神经毒剂类似物的水解

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-10 DOI: 10.1016/j.bej.2025.109862

Priyamedha Yadav, Manik Goel, Rinkoo Devi Gupta

{"title":"Rational designing of human senescence marker protein 30 for enhanced hydrolysis of V-type nerve agent analog","authors":"Priyamedha Yadav, Manik Goel, Rinkoo Devi Gupta","doi":"10.1016/j.bej.2025.109862","DOIUrl":"10.1016/j.bej.2025.109862","url":null,"abstract":"<div><div>Organophosphate (OP) nerve agents are a class of compounds known for high toxicity and rapid action, posing a significant threat to human health. Catalytic bio-scavengers have gained substantial attention due to the limited protective efficacy and adverse side effects of chemical drugs in treating OP poisoning. However, the low catalytic efficiency remains a key limitation to their effectiveness and broader applications. Unlike existing human catalytic bio-scavengers such as huPON1, which lack activity against V-type nerve agents, the human Senescence Marker protein 30 (huSMP30) can detoxify a surrogate V-type nerve agent, i.e., Demeton-S; however, its hydrolytic activity is too low to be utilized as a drug. Therefore, in this study, the huSMP30 has been engineered through a rational protein designing approach to enhance its catalytic activity for Demeton-S. Initially, <em>in silico</em> analysis identified six key amino acid residues (E18, A62, K106, N154, D157, and D204), which were targeted for mutational analysis. Fourteen mutants, including single, double, and triple mutants, were created by site-directed mutagenesis, recombinantly produced, and analyzed <em>in vitro</em>. Substitution mutation at position 106 significantly enhanced the catalytic efficiency (<em>kcat/Km</em>) by up to ∼26-fold. In conclusion, the engineered huSMP30 may act as an effective catalytic bio-scavenger for the hydrolysis of Demeton-S; therefore, it could be utilized for the treatment of V-type nerve agent poisoning.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"223 ","pages":"Article 109862"},"PeriodicalIF":3.7,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144631952","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Optimal porosity in zeolitic imidazole framework for cost-effective furfural removal in biomass hydrolysates 沸石型咪唑骨架的最佳孔隙度对生物质水解液中糠醛的经济高效去除

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-09 DOI: 10.1016/j.bej.2025.109858

Kanghong Wang , Chaozhong Xu , Wei Xiong, Shanshan Tong, Jia Ouyang, Xiaoli Gu

{"title":"Optimal porosity in zeolitic imidazole framework for cost-effective furfural removal in biomass hydrolysates","authors":"Kanghong Wang , Chaozhong Xu , Wei Xiong, Shanshan Tong, Jia Ouyang, Xiaoli Gu","doi":"10.1016/j.bej.2025.109858","DOIUrl":"10.1016/j.bej.2025.109858","url":null,"abstract":"<div><div>Efficient removal of fermentation inhibitors, particularly furan aldehydes like furfural is crucial for enhancing microbial fermentation efficiency in lignocellulosic biomass processing. In this study, zeolitic imidazolate framework-8 with different molar ratios were synthesized by regulating the Zn<sup>2 +</sup> /2-methylimidazole molar ratio (from 1:4–1:15), and their structural properties and adsorption performance were systematically evaluated. The results showed that the pore structure was crucial to the adsorption performance, and the pore structure was optimal when the Zn<sup>2+</sup>/2-methylimidazole ratio was 1:8. The optimized zeolitic imidazole framework-8 showed rapid, selective and efficient adsorption of furfural under different operating conditions, thereby preventing the formation of inhibitors from affecting fermentation while minimizing sugar loss. This study provides a generalizable design principle for the development of adsorbents in biomass conversion. This work provides a scalable solution for biomass hydrolysate detoxification, with potential cost advantages due to the material’s reusability and high adsorption capacity compared to conventional resin-based methods.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"222 ","pages":"Article 109858"},"PeriodicalIF":3.7,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144597066","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Zinc extraction from cast-house dust using the chemolithotrophic microorganism Acidithiobacillus thiooxidans 利用化化嗜硫微生物酸化硫杆菌从铸造厂粉尘中提取锌

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-08 DOI: 10.1016/j.bej.2025.109859

Rebeka Frueholz , Clemens Habermaier , Sabine Spiess , Ludwig Birklbauer , Georg M. Guebitz , Marianne Haberbauer

{"title":"Zinc extraction from cast-house dust using the chemolithotrophic microorganism Acidithiobacillus thiooxidans","authors":"Rebeka Frueholz , Clemens Habermaier , Sabine Spiess , Ludwig Birklbauer , Georg M. Guebitz , Marianne Haberbauer","doi":"10.1016/j.bej.2025.109859","DOIUrl":"10.1016/j.bej.2025.109859","url":null,"abstract":"<div><div>The continuous generation of steel by-products calls for innovative recycling solutions in order to strive towards a circular economy. In this study, the bioleaching potential of the chemolithotrophic microorganism <em>A. thiooxidans</em> was evaluated to extract unwanted Zn from the steel by-product cast-house dust (CHD) while specifically retaining Fe within the dust matrix to enable on-site recycling. A fed-batch stirred tank reactor was employed, and process stability was assessed through a series of replicates at a CHD concentration of 125 g L⁻¹ . Two distinct aerations 0.42 vvm and 0.50 vvm were examined during the course of this investigation. After 14 days, a maximum of 75 ± 5 % Zn underwent solubilization while only 9 ± 2 % of Fe was solubilized. However, after 7 days the majority of Zn was extracted resulting in an average leaching efficiency of 68 ± 2 % for Zn and 5 ± 2 % for Fe. Although increasing the aeration rate did not significantly improve the <em>k<sub>L</sub>a</em> value, it slightly enhanced O₂ availability, highlighting the need for further optimization of the aeration setup.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"222 ","pages":"Article 109859"},"PeriodicalIF":3.7,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144597067","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Microbial community analysis in microbial fuel cell under non-strict anaerobic condition: synchronous generation of power and tobacco wastewater treatment 非严格厌氧条件下微生物燃料电池微生物群落分析：同步发电与烟草废水处理

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-08 DOI: 10.1016/j.bej.2025.109857

Yutong Liu , Cong Chen , Suxuan Li , Shuhe Ma , Xiaoyu Chen , Jian Hong , Xiaohu Li , Zhiyong Wu

{"title":"Microbial community analysis in microbial fuel cell under non-strict anaerobic condition: synchronous generation of power and tobacco wastewater treatment","authors":"Yutong Liu , Cong Chen , Suxuan Li , Shuhe Ma , Xiaoyu Chen , Jian Hong , Xiaohu Li , Zhiyong Wu","doi":"10.1016/j.bej.2025.109857","DOIUrl":"10.1016/j.bej.2025.109857","url":null,"abstract":"<div><div>Tobacco wastewater poses a significant environmental threat due to its toxic constituents, which are difficult to remove using conventional treatment methods. Microbial fuel cells (MFCs) are promising devices harnessing the catabolic activity of microorganisms to produce electricity with wastewater treatment. In this study, MFCs were constructed to evaluate the performance of tobacco wastewater treatment and electrical energy generation under mildly anaerobic conditions. The MFCs supplied with various additional carbon sources (e.g., acetate, glucose, propionate, and butyrate) exhibited different acclimation capacities for tobacco wastewater treatment. The maximum voltage output of 0.58 V and power density of 193.60 mW/m<sup>2</sup> were obtained, along with COD removal of 73.4 ± 2.93 %. The microbial community compositions revealed that <em>Trichococcus</em> was highly abundant in the anodic biofilm enrichment stage (7.46 %-21.07 %) and the wastewater treatment stage (16.22 %-36.55 %), which may play an important role in the electrical energy generation. This study firstly demonstrates that MFC is an effective and environmentally friendly technology for tobacco wastewater treatment and electrical energy generation under non-strict anaerobic condition.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"222 ","pages":"Article 109857"},"PeriodicalIF":3.7,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144580211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Combined strategies to improve the heterologous expression of glucose oxidase from Aspergillus cristatus in Komagataella phaffii 提高法菲黑马霉葡萄糖氧化酶异源表达的联合策略

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-05 DOI: 10.1016/j.bej.2025.109856

Qingzhu Wang , Lulu Chen , Jia Zheng , Bowen Ke , Yi’an Chen , Yaping Zhang , Shuangyan Han

{"title":"Combined strategies to improve the heterologous expression of glucose oxidase from Aspergillus cristatus in Komagataella phaffii","authors":"Qingzhu Wang , Lulu Chen , Jia Zheng , Bowen Ke , Yi’an Chen , Yaping Zhang , Shuangyan Han","doi":"10.1016/j.bej.2025.109856","DOIUrl":"10.1016/j.bej.2025.109856","url":null,"abstract":"<div><div>Glucose oxidase (EC 1.1.3.4, GOD) is a key industrial enzyme with broad applications across the food, pharmaceutical, clinical, and chemical industries. In this study, a glucose oxidase, designated <em>c</em>GOD, was identified from <em>Aspergillus cristatus</em> and heterologously expressed in <em>Komagataella phaffii.</em> Biochemical characterization revealed that <em>c</em>GOD exhibited optimal activity at 40 °C and pH 6.0. To enhance its production, a combinatorial strategy was implemented, involving optimization of promoter and signal peptide, amplification of gene copy number, and modulation of the secretory pathway. As a result, extracellular <em>c</em>GOD activity increased to 967.23 U/mL in shake flask culture. Notably, in a 15 L fed-batch fermenter, the <em>c</em>GOD enzyme activity reached 11,655 U/mL, surpassing previously reported GOD activity levels. The successful identification and efficient expression of <em>c</em>GOD from <em>A. cristatus</em> provide valuable insights for the development of high-yield microbial strains for industrial enzyme production.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"224 ","pages":"Article 109856"},"PeriodicalIF":3.7,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144890788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Layered modification for enhancing O-Succinyl-L-homoserine biosynthesis with a non-auxotrophic probiotic Escherichia coli Nissle 1917 非营养不良益生菌大肠杆菌Nissle 1917增强o -琥珀酰- l-同丝氨酸生物合成的层状修饰

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-04 DOI: 10.1016/j.bej.2025.109852

Kun Niu , Yao-Yao Wang , Si-Min Huang , Ming-Quan Liao , Zhi-Qiang Liu , Yu-Guo Zheng

{"title":"Layered modification for enhancing O-Succinyl-L-homoserine biosynthesis with a non-auxotrophic probiotic Escherichia coli Nissle 1917","authors":"Kun Niu , Yao-Yao Wang , Si-Min Huang , Ming-Quan Liao , Zhi-Qiang Liu , Yu-Guo Zheng","doi":"10.1016/j.bej.2025.109852","DOIUrl":"10.1016/j.bej.2025.109852","url":null,"abstract":"<div><div>O-Succinyl-L-homoserine (OSH) is a promising platform compound with extensive application prospects in medicine, feed, agriculture, and food. Many scholars have dedicated significant efforts to the rational engineering of microbial cell factories, particularly in <em>Escherichia coli</em>, aiming to achieve efficient production of OSH. In this study, modular genetic engineering was performed to achieve layered modification of the OSH biosynthesis pathway. Firstly, an OSH producing chassis strain was constructed based on the wild-type probiotic <em>E. coli</em> Nissle 1917 (EcN) via knocking out the negative genes, dynamically regulating the synthesis of by-product, relieving the feedback inhibition of key enzymes, and weakening the degradation pathway. Furthermore, the supply of the key precursors L-homoserine, L-aspartate and oxaloacetate was strengthened to drive the carbon flux into the OSH biosynthesis. In addition, the regeneration of cofactor NADPH further increased the OSH titer to 13.44 ± 0.77 g/L in the shake flask. Fed-batch fermentation in the 5-L bioreactor showed strain OSHY33 produced 80.79 ± 2.10 g/L of OSH within 44 h, representing the highest OSH productivity of 1.84 g/L/h to date. This research has laid a solid foundation for OSH production through microbial fermentation.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"222 ","pages":"Article 109852"},"PeriodicalIF":3.7,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144570771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 novel bioengineered spider silk-based particle for the delivery system of chemotherapy drugs 一种新型生物工程蜘蛛丝为基础的化疗药物递送系统颗粒

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-03 DOI: 10.1016/j.bej.2025.109855

Yongqiang Zhou , Lanxiao Shen , Zhimin Zheng , Mingrou Guo , Zheng Peng

{"title":"A novel bioengineered spider silk-based particle for the delivery system of chemotherapy drugs","authors":"Yongqiang Zhou , Lanxiao Shen , Zhimin Zheng , Mingrou Guo , Zheng Peng","doi":"10.1016/j.bej.2025.109855","DOIUrl":"10.1016/j.bej.2025.109855","url":null,"abstract":"<div><div>Spider silk protein (spidroin) is the primary constituent of spider silk fibers and exhibits remarkable biocompatibility, thermal stability, and biodegradability, making it a promising biomaterial for biomedical applications, particularly in drug delivery. Currently, drug delivery systems utilizing spidroins predominantly focus on engineered dragline spidroins. However, research on other spidroin types—such as aciniform spidroins (AcSp) —in their application as biomaterials within drug delivery systems remains very limited. Here, we engineered the recombinant spidroin AS2, derived from the <em>Araneus ventricosus</em> AcSp2 protein repeat domain, and designed a modified version, eAS2, by incorporating an additional glutamic acid residue. Both AS2 and eAS2 proteins exhibited self-assembly properties, forming non-toxic nanoparticles with similar secondary structure content in potassium phosphate buffer but differed in zeta potential values. Although eAS2 particles demonstrated high loading efficiency for positively charged drugs like mitoxantrone (MTX), a rapid release rate was also noted for MTX-loaded eAS2 particles. Conversely, the combination of efficient loading and a desirable slow release profile renders eAS2 particles an optimal delivery system for neutral and positive charged drugs, such as etoposide (ETP) and doxorubicin (DOX).</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"222 ","pages":"Article 109855"},"PeriodicalIF":3.7,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144557403","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Chalcone isomerase-like (CHIL) impedes the lactone shunt and modulates flux partitioning in isoflavonoid biosynthesis 查尔酮异构酶样（CHIL）阻碍内酯分流并调节异黄酮生物合成中的通量分配

IF 3.7 3区生物学

Biochemical Engineering Journal Pub Date : 2025-07-03 DOI: 10.1016/j.bej.2025.109853

Lee Marie Raytek , Brandon Corey Saltzman , Meha Sharma , Soon Goo Lee , Mehran Dastmalchi

{"title":"Chalcone isomerase-like (CHIL) impedes the lactone shunt and modulates flux partitioning in isoflavonoid biosynthesis","authors":"Lee Marie Raytek , Brandon Corey Saltzman , Meha Sharma , Soon Goo Lee , Mehran Dastmalchi","doi":"10.1016/j.bej.2025.109853","DOIUrl":"10.1016/j.bej.2025.109853","url":null,"abstract":"<div><div>The reconstitution of biosynthetic pathways in heterologous hosts is often challenged by the transition to a foreign cellular environment, lacking compatible structural or regulatory features. Auxiliary or non-catalytic proteins can play a critical role in guiding metabolic flux. Chalcone isomerase-like (CHIL) is a non-catalytic protein known to serve as a partner to chalcone synthase (CHS) in flavonoid biosynthesis, rectifying its promiscuous activity and preventing the formation of by-products, such as <em>p</em>-coumaroyltriacetic acid lactone (CTAL). Here, we extended the functional analysis of CHILs to the legume-characteristic isoflavonoid pathway. We assessed CHIL orthologs, using sequence analysis and structural modelling to predict their respective binding capacities, followed by functional characterization. The addition of CHIL to enzyme assays containing CHS, alone or with downstream enzymes, chalcone reductase (CHR) and chalcone isomerase (CHI), reduced CTAL levels (up to 42 %) while simultaneously increasing the output of desired intermediates. Combinatorial yeast biotransformation assays revealed that CHIL plays a crucial role in directing metabolic flux through chalcone and flavanone pathways, and, with co-expression of isoflavone synthase (IFS), in isoflavone biosynthesis. The inclusion of CHIL in engineered yeast strains enhanced overall titers and, unexpectedly, promoted partitioning toward the deoxy-branch (isoliquiritigenin, liquiritigenin, and daidzein) up to 50 %, with a 33 % increase in final daidzein titers. Therefore, we have revealed an expanded role for CHIL as an auxiliary protein in isoflavonoid biosynthesis and underscored the utility of non-catalytic proteins in metabolic engineering.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"222 ","pages":"Article 109853"},"PeriodicalIF":3.7,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144580204","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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