Journal of Biological Engineering最新文献

{"title":"Application of extracellular vesicles in the CRISPR-based diagnosis and treatment: possibilities and challenges.","authors":"Eunhye Cho, Jehyung Lee, Jiyu Kim, Jaewon Choi, Mikyung Kang, Jayeon Song","doi":"10.1186/s13036-026-00673-6","DOIUrl":"10.1186/s13036-026-00673-6","url":null,"abstract":"<p><p>The CRISPR–Cas system has revolutionized molecular diagnostics and gene editing, yet its clinical translation is hindered by delivery barriers, off-target activity, immunogenicity, and manufacturing challenges. Compared with viral vectors and synthetic non-viral carriers such as lipid nanoparticles, extracellular vesicles (EVs) offer a biologically derived delivery platform with superior biocompatibility, reduced immunogenicity, intrinsic cargo protection, and natural barrier-crossing capability. Engineered EVs can further achieve cell- or tissue-specific targeting. In diagnostics, endogenous EV proteins and nucleic acids provide disease-informative signatures that can interface with CRISPR nuclease readouts for highly sensitive detection. This review summarizes the therapeutic and diagnostic potential of EV-CRISPR platforms, covering strategies for loading CRISPR cargos (producer-cell engineering, post-isolation methods), cargo formats, and surface targeting approaches. We evaluate preclinical performance with attention to biodistribution, safety, innate and adaptive immune responses, and genomic integrity, as well as analytical assays and scalable manufacturing considerations essential for clinical translation. Finally, we discuss emerging opportunities, including AI-guided optimization of EV-CRISPR design and integrated EV platforms that combine disease detection with therapeutic intervention, highlighting their promise for advancing precision medicine.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13202827/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147698875","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multi-technique characterization of gold nanoparticle-based DNA biosensor with smartphone-enabled photometry for accelerated detection of Campylobacter jejuni.","authors":"Anthony James Franco, Finnian James, Leah Wilson, Woubit Abebe, Tina Conklin, Roger Stearns, Mollie Woods, Evangelyn Alocilja","doi":"10.1186/s13036-026-00677-2","DOIUrl":"10.1186/s13036-026-00677-2","url":null,"abstract":"<p><p>BACKGROUND: Campylobacter is one of the most infectious foodborne pathogens, often associated with consuming contaminated poultry products. Many Campylobacter infections are not reported until the symptoms become severe, making it necessary to detect them rapidly in food production systems as part of a preventative intervention strategy. In this study, a gold nanoparticle (GNP)-based DNA biosensor that utilizes digital photometric signals from smartphone images was developed to rapidly detect Campylobacter jejuni in drainage water and poultry rinse samples. RESULTS: Spectrophotometric measurements, hydrodynamic size, zeta potential, and dark-field microscopy confirm that hybrids were successfully formed between GNP, oligonucleotide probe, and C. jejuni DNA using an in-situ approach. Triggering GNP aggregation upon hybridization, the nano-biosensor signals obtained through smartphone-enabled digital photometry were strongly correlated with spectrophotometry. Based on the smartphone-enabled signal, the nano-biosensor demonstrated a high degree of selectivity for C. jejuni and detected 103 and 106 colony-forming units (CFU)/mL in unenriched drainage water and poultry rinse samples, respectively. CONCLUSIONS: This work demonstrates a GNP-based DNA biosensor employing cadF-targeted hybridization and smartphone-enabled digital photometry for the detection of Campylobacter jejuni in real samples. Diffusion-limited aggregation of unhybridized GNPs yields distinct optical responses, enabling the detection of target DNA in drainage water and poultry rinse. These findings provide a foundation for future validation studies in samples critical to monitoring foodborne pathogens across various food production and processing systems. Moreover, they highlight the potential of the smartphone-aided nano-biosensing system as a cost-effective, field-deployable means of pathogen surveillance. </p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13238093/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147662445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microfluidics-based cell recognition through optimizing suspended cell staining techniques and artificial intelligence.","authors":"Areen K Al-Bashir, Lamis R Bany Issa, Haneen Ababneh, Farah Alshami, Esra'a Al-Shawwa, Ala'a Al-Rashdan, Ruba E Khnouf","doi":"10.1186/s13036-026-00678-1","DOIUrl":"10.1186/s13036-026-00678-1","url":null,"abstract":"<p><p>PURPOSE: Blood staining is necessary to differentiate between its various cells. It is typically performed on a microscopic slide that contains a blood film. However, in some applications, there is a need to stain blood in suspension. METHODS: Three techniques were optimized (Romanowsky stains, Hematoxylin stain, and our proposed modified Romanowsky stain) for cell staining before injection into a microfluidic device. Images of stained cells in the microfluidic device were captured under an optical microscope. The effectiveness of staining techniques in cell differentiation was evaluated using color-based k-means clustering. Then, four pre-trained models (Alexnet, VGG16, Resnet, and Densnet) were applied to extract features and classify RBCs, WBCs, and CTCs. RESULTS: Only images showing clear cell borders were retained. The models achieved high testing accuracy, reaching 100%, 98%, 98%, and 95.34%, respectively. CONCLUSIONS: This high accuracy was achieved without the use of fluorescence labeling or expensive materials, thereby preserving the cells alive. Based on these results, the proposed staining technique was the most effective in differentiating between WBCs, RBCs, and CTCs, where all WBCs and RBCs were classified correctly. </p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13185322/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147654273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fabrication and characterization of 3D-printed polyacrylonitrile scaffolds for the neural differentiation of mesenchymal stem cells via exosomes.","authors":"Elham Hoveizi, Mostafa Sayahi","doi":"10.1186/s13036-026-00648-7","DOIUrl":"10.1186/s13036-026-00648-7","url":null,"abstract":"<p><p>Exosomes mediate signaling that guides differentiation by delivering factors that influence cellular fate and gene pathways. Three-dimensional (3D)–printed scaffolds provide customizable matrices that create precise microenvironments for differentiation and can synergize with exosome cues to enhance lineage specification and tissue regeneration. This study evaluated the neurogenic differentiation potential of exosomes derived from neural stem cells (NSCs-exs) on adipose-derived mesenchymal stem cells (ADMSCs) cultured on a polyacrylonitrile (PAN) scaffold prepared by 3D printing. NSCs-exs were characterized by transmission electron microscopy (TEM), Western blotting, and dynamic light scattering (DLS). The PAN scaffold was fabricated by a solution-printing method using a 3D printer to deposit a 10% PAN polymer mixture. ADMSCs were cultured on the scaffold and treated with 10 µg/mL NSCs-exs for 14 days. Differentiation was assessed by examining the expression of Nestin, Map2, Tuj-1, and NF using immunocytochemistry and RT-PCR. NSCs-exs were characterized by a diameter of 44.79 nm, a lipid bilayer structure, and expression of the markers CD81, CD9, and CD63, with a total protein concentration of 160 µg/mL. Microstructural analysis of the PAN scaffold revealed a homogeneous and organized architecture with an average fiber diameter of approximately 470 nm. The scaffold exhibited appropriate chemical bonding, sufficient mechanical strength, and biocompatibility. A significant increase (P ≤ 0.05) in neural-marker expression indicated successful neural differentiation of ADMSCs under NSCs-exs treatment in 3D culture. Exosome-guided differentiation on 3D PAN scaffolds promotes neural lineage commitment, reinforcing exosomes as reparative tools for neural regeneration.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13173745/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147615805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Residual-hybrid dynamic response-surface/machine-learning framework for predicting malachite green removal and biodegradation by a bacterial consortium.","authors":"Ebtehag A E Sakr, Marwa S Hassan, Mervat El-Sedik, Manal M Rekaby, Ghada M El-Sayed","doi":"10.1186/s13036-026-00663-8","DOIUrl":"10.1186/s13036-026-00663-8","url":null,"abstract":"<p><p>BACKGROUND: The discharge of dye-containing effluents is a global environmental concern, particularly for synthetic cationic dyes such as malachite green (MG). This study aimed to develop and experimentally validate a hybrid dynamic response-surface/machine-learning framework, coupled with an optimized bacterial consortium, for predicting and enhancing MG removal and elucidating its biodegradation pathway. RESULTS: A four-strain comprising Azotobacter chroococcum, Azotobacter salinestris, Stenotrophomonas maltophilia, and Sphingomonas kyeonggiensis was first optimized by mixture design, where an appropriate proportion of A. chroococcum and S. maltophilia achieved 83.51% MG decolorization. A Box–Behnken design (BBD) was then applied to optimize initial MG concentration, inoculum size, and incubation time; the response-surface model predicted a maximum removal of 96.56% at 100 mg/L, 7% inoculum, and 72 h, in good agreement with the experimental data. The resulting RSM-based dynamic response-surface model used as a baseline, and residual machine-learning regressors were trained on experimentally derived residuals (experimental MG removal minus baseline prediction). Among standalone learners, CatBoost provided the best performance on the exploratory 80/20 train–test split, whereas under leave-one-out cross-validation, the hybrid residual model (dynamic response-surface + Ridge regression) emerged as the most reliable predictor, yielding closer agreement with the experimental MG removal than the baseline model. Biodegradation of MG by the optimized consortium was experimentally validated by Fourier-transform infrared (FTIR) spectroscopy, liquid chromatography–mass spectrometry (LC–MS) and UV–visible spectroscopy, which confirmed disruption of the chromophoric structure and formation of less toxic metabolites. Toxicity assay indicated that the degradation products were non-toxic towards the tested pathogenic bacteria. ISSR molecular typing revealed that primer ISSR-12 exhibited strong discriminatory power, generating nine strain-specific bands, whereas primer ISSR-1 produced a higher proportion of monomorphic bands. CONCLUSIONS: This work introduces a small-sample, hybrid dynamic response-surface/machine-learning framework, experimentally validated against MG biodegradation experiments, which integrates an RSM-based baseline as an explicit feature in residual learning. Under leave-one-out cross-validation (N = 17), the RSM baseline achieved R² = 0.988 and RMSE = 2.62% MG removal, while the best hybrid residual model (dynamic response-surface + Ridge regression) yielded R² = 0.987 and RMSE = 2.79%, confirming the robustness of the proposed framework. </p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13063629/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147609069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Precision Fc remodeling via glycosylation-competent CHO display enables ultra-selective FcγRIIIa targeting and enhanced antitumor activity.","authors":"Migyeong Jo, Suyeon Kim, Sanghwan Ko, Munsu Kyung, Seunghyeon Lee, Woo Hyung Ko, Wonju Lee, Sang Taek Jung","doi":"10.1186/s13036-026-00671-8","DOIUrl":"10.1186/s13036-026-00671-8","url":null,"abstract":"<p><p>BACKGROUND: Improving tumor cell clearance by therapeutic antibodies remains a translational bottleneck because wild-type IgG1 Fc typically elicits suboptimal NK cell–mediated antibody-dependent cellular cytotoxicity (ADCC), necessitating Fc engineering to enhance activating FcγR engagement while preserving antigen specificity and manufacturability. FcγRIIIa (CD16A) is the principal activating receptor on NK cells, and its clinical relevance is underscored by the FCGR3A-158V/F polymorphism, which modulates IgG1 Fc affinity and therapeutic response. Because FcγRIIIa engagement critically depends on Fc glycosylation, microbial display platforms lacking mammalian glycan processing are limited in capturing Fc–FcγRIIIa energetics and selectivity. RESULTS: We developed a glycosylation-integrated Fc engineering platform using CHO surface display to screen glycosylated Fc libraries in a post-translationally accurate context. Multiparameter flow-cytometric selection with FcγRIIIa binding and FcγRIIb counter-screening enabled iterative isolation of PS-series Fc variants with ultra-selective, allotype-compatible FcγRIIIa recognition. Lead variants achieved 262-fold (158V) and 497-fold (158F) FcγRIIIa affinity gains while reducing FcγRIIb binding by up to 4.2-fold, expanding activating-to-inhibitory selectivity up to 2,096. This exceeded clinically deployed FcγRIIIa-enhancing Fc variants by 525-fold and 108-fold, respectively, in activating-to-inhibitory selectivity, compared with DE (S239D/I332E; used in tafasitamab) and VLPLL (L235V/F243L/R292P/Y300L/P396L; used in margetuximab). When grafted onto trastuzumab, PS variants enhanced NK cytotoxicity and improved tumor control in a trastuzumab-refractory xenograft model. Modular transfer to cetuximab and rituximab also increased cytotoxic activity. CONCLUSIONS: This study establishes a glycosylation-integrated CHO display platform for precision FcγRIIIa-targeted Fc engineering, generating modular effector domains with broad allotype compatibility, minimal FcγRIIb binding, and robust therapeutic potential in cancer immunotherapy. </p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13170290/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147592351","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The non-scaffold effects of natural polysaccharides in promoting organ regeneration: a review.","authors":"Sheng-Ping Lei, Pan Chang, Li Li, Yan Niu, Jun Yu, Jian-Li Gao","doi":"10.1186/s13036-026-00655-8","DOIUrl":"10.1186/s13036-026-00655-8","url":null,"abstract":"<p><p>Natural polysaccharides are known for their scaffold-based roles in tissue engineering, but their non-scaffold bioactivities in promoting organ regeneration remain poorly understood.This review aims to systematically summarizes the latest research advances in natural polysaccharides promoting organ regeneration through non-scaffold mechanisms, exploring their multifaceted actions, including creating a pro-regenerative microenvironment (e.g., immune regulation, extracellular matrix (ECM) remodeling, and gut microbiota-mediated short-chain fatty acids (SCFAs)), regulating stem cell proliferation and differentiation, promoting angiogenesis, and other effects like epigenetic regulation, antioxidant activity, and mitochondrial function modulation. Also outline their broad application potential in the regeneration and repair of multiple organ systems, including skin, heart, nerves, liver, and intestine. Furthermore, we discuss current challenges and future development directions, aiming to effectively exploit the therapeutic potential of natural polysaccharides in regenerative medicine and provide novel strategies for addressing organ repair challenges.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13162400/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147592381","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Balancing stromal-induced complexity in 3D ovarian cancer models through heterotypic co-culture with fibroblasts or architected micro-scaffolds.","authors":"Eglė Žymantaitė, Karolina Limanovskaja, Linas Jonušauskas, Vita Pašukonienė, Neringa Dobrovolskienė, Agata Mlynska","doi":"10.1186/s13036-026-00670-9","DOIUrl":"10.1186/s13036-026-00670-9","url":null,"abstract":"<p><p>Tumor heterogeneity limits the reproducibility and interpretability of in vitro ovarian cancer models. In this study, we established a comparative framework that intentionally modulates stromal-induced complexity through two complementary approaches: scaffold-free stromal co-culture with patient-derived cancer-associated fibroblasts (CAFs) or fibroblast cell line WS1, and scaffold-based micro-scaffolds printed using two-photon polymerization (2PP) with poly(ethylene glycol) diacrylate (PEGDA). We analyzed four ovarian cancer cell lines (A2780, SKOV3, COV362 and OV7,) stemness/epithelial-mesenchymal transition (EMT), and TGF-β. We then evaluated their 3D spheroid formation and growth dynamics across different seeding densities. In ultra-low-attachment plates, SKOV3, COV362, and OV7 formed round, compact spheroids across different seeding densities, while A2780 produced loose aggregates. Co-culturing with early-passage CAFs or WS1 fibroblast cell line (in 2:1, 1:1, and 1:2 ratios of cancer cells : fibroblast) rescued A2780‘s ability to form spheroids and increased the compactness of OV7 and COV362 in a line- and ratio-dependent manner in co-culture with CAFs. Compared to 2D cultures, 3D spheroids exhibited higher expression of stemness/EMT and angiogenesis related genes. In contrast, the PEGDA scaffolds (with pore sizes of 65, 100, and 130 µm) standardized early attachment and surface coverage for both spheroid-competent line SKOV3, and a line that does not typically form spheroids, A2780. These scaffolds also shifted EMT/stemness gene expression without elevating vascular endothelial growth factor (VEGF) levels compared to scaffold-free 3D cultures. Together, these results provide practical guidance: utilize fibroblast co-culture to study the stromal-induced complexity and cell-to-cell interactions, including the rescue of cell lines that typically do not form spheroids, and employ printed micro-scaffolds to control 3D culture variability.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13159347/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147581241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioengineered curcumin-SCFA co-loaded PLGA nanoparticles for modulating inflammation and epithelial barrier integrity in radiation-induced enteritis.","authors":"Kun He, Chengyan Zhang, Yuan Li, Xiuxiu Li, Xin Zhang","doi":"10.1186/s13036-026-00645-w","DOIUrl":"10.1186/s13036-026-00645-w","url":null,"abstract":"<p><p>Radiation enteritis (RE) is a common side effect of abdominal radiotherapy, which causes intestinal inflammation and impairment of epithelial barrier. In this study, we developed a bioengineered dual-delivery nanosystem by encapsulating curcumin and microbiota-derived short-chain fatty acids (SCFAs: acetate, propionate, and butyrate) into PLGA nanoparticles (Cur-PLGA NPs@SCFAs). The formulation encapsulated 66% of curcumin and released it slowly over 48 h, with about 72.6% of it being released at pH 7.4. In vitro studies using Caco-2 cells, the Cur-PLGA NPs@SCFAs maintained good cytocompatibility, with cell viability over 81.5% at 40 µg/mL as well as reduced intracellular reactive oxygen species to 65.1% of control levels. Wound closure reached 86.2% at 48 h whereas compared to untreated cells (42.7%). In vivo mice model, high-dose administration (40 mg/kg) markedly improved disease outcomes, reducing the Disease Activity Index to 3.8 ± 0.4 compared with 8.6 ± 0.5 in the RE group, while restoring colon length (7.1 ± 0.2 cm) and spleen index (0.29 ± 0.01%). Pro-inflammatory mediators IL-6 and TNF-α were decreased, accompanied by suppression of NF-κB p65 signaling, whereas IL-10 and the tight junction proteins ZO-1 and occludin were increased. Antioxidant defenses (GSH, SOD, and T-AOC) were recovered, and cecal SCFA levels were nearly doubled, collectively indicating a protective effect on intestinal inflammation and barrier integrity.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13151141/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147573806","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pathway reconstruction of Corynebacterium glutamicum for C30 carotenoid 4,4'-diaponeurosporene production.","authors":"Chi Young Hwang, Eui-Sang Cho, Seong-Bo Kim, Myung-Ji Seo","doi":"10.1186/s13036-026-00674-5","DOIUrl":"10.1186/s13036-026-00674-5","url":null,"abstract":"<p><p>4,4′-Diaponeurosporene (4,4′-DIA) is a rare C30 carotenoid with strong antioxidant and immune-enhancing properties. However, microbial production of 4,4′-DIA has been limited by low titers and the simultaneous formation of undesired carotenoid byproducts. In this study, Corynebacterium glutamicum was rationally engineered to enable selective 4,4′-DIA biosynthesis through pathway reconstruction. To redirect metabolic flux toward C30 carotenoids, the native C50 carotenoid biosynthetic gene cluster was deleted and the precursor supply was reconfigured by replacing the core C20 derivative pathway with ispA to produce C15 precursor. A gene module enabling high-level and selective production of 4,4′-DIA was established through the combinatorial expression of crtM and crtN in a constitutive vector system. Pathway analysis predicted crtN to be a bottleneck in 4,4′-DIA biosynthesis, and its expression was strengthened through promoter introduction, leading to improved production level. Deletion of poxB and ldhA combined with ribK overexpression enhanced both 4,4′-DIA biosynthetic gene expression and production, likely due to improvement carbon flux and cofactor supply. The final engineered strain was successfully transferred to a 5-L fed-batch fermentation with a 4,4′-DIA titer of 656.3 mg/L without any inducer. This work demonstrates the selective production of the rare and non-commercial C30 carotenoid 4,4′-DIA in C. glutamicum, establishing it as a robust platform for diversified carotenoid biosynthesis.</p>","PeriodicalId":15053,"journal":{"name":"Journal of Biological Engineering","volume":" ","pages":""},"PeriodicalIF":6.3,"publicationDate":"2026-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13151236/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147573793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}