Biotechnology and Bioengineering最新文献

{"title":"Cast‐Molded Channelized Hydrogel Scaffolds With Stereolithography‐Printed Templates","authors":"Chi Wang, Yingge Zhou","doi":"10.1002/bit.70007","DOIUrl":"https://doi.org/10.1002/bit.70007","url":null,"abstract":"Creating internal vascular networks within hydrogel scaffolds is crucial for providing the encapsulated cells with the necessary nutrients, oxygen, and metabolic exchange. Current methods for hydrogel scaffold fabrication face significant hurdles, including the challenge of forming sufficient internal channels, achieving precise scaffold geometry, and maintaining high cell viability, often compromised by the fabrication process and properties of the polymer materials used. Stereolithography (SLA) emerges as a promising 3D printing technique due to its exceptional precision, efficiency, and resolution, allowing for the creation of complex geometries with fine detail. This paper explores the application of SLA as a novel strategy to fabricate hydrogel scaffolds with interconnected small diameter channels, surpassing the capabilities of fused deposition modeling method to create templates. The encapsulated fibroblasts grown in the hydrogel scaffold containing channels showed significantly elevated cell viability compared to the ones without any channels. The capability of this SLA‐assisted strategy to create channel structures with encapsulated cells demonstrate significant potential for generating 3D artificial tissue composites with precisely controlled micron‐scale channels.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"62 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144515242","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}

{"title":"Modular RNAi Pathway Engineering Enhances Plasmid Copy Number Control in Yeast Bioproduction System","authors":"Qianru Cai, Manman Wang, Jinmei Zhu, Bangce Ye, Xiaohe Chu, Jiequn Wu","doi":"10.1002/bit.70014","DOIUrl":"https://doi.org/10.1002/bit.70014","url":null,"abstract":"Rational design of microbial cell factories requires precise coordination of gene dosage and expression dynamics to optimize metabolic flux while minimizing cellular burden. In this study, an unexpected plasmid copy number amplification was identified following the reconstruction of an orthogonal RNA interference (RNAi) system in <jats:italic>Saccharomyces cerevisiae</jats:italic>, and a synthetic biology chassis with dynamically programmable plasmid copy numbers was developed. By integrating heterologous RNAi pathway genes from <jats:italic>Saccharomyces castellii</jats:italic> and designing sequence‐specific small interfering RNAs (siRNAs) targeting plasmid‐encoded selection markers, we established a chemically inducible gene dosage control platform capable of achieving plasmid copy number amplification up to 7.13‐fold. Application of this RNAi‐mediated copy number modulation to the carotenoid biosynthetic pathway resulted in an 18.6‐fold increase in lycopene titers compared to static plasmid systems. This study presents an innovative approach for dynamic plasmid copy number regulation in <jats:italic>S. cerevisiae</jats:italic> to enable high‐efficiency gene dosage control, further enriching the toolkit for synthetic metabolic regulation. This strategy exhibits significant potential for enhancing the production performance of microbial cell factories and offers novel perspectives for metabolic engineering optimizations within the synthetic biology framework.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"47 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144515212","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}

{"title":"Enhancement of Chondrogenesis by a Microcavitary Hydrogel Coculture System Synergizing With SDF‐1α and TGF‐Β3","authors":"Qian Pan, Hui Gao, Weixian Su, Yupeng Nie, Rixu Liu, Weiqiang Dong, Yongchang Yao","doi":"10.1002/bit.70008","DOIUrl":"https://doi.org/10.1002/bit.70008","url":null,"abstract":"A favorable microenvironment is of great significance for the repair of cartilage injury. In our previous study, a coculture system was devised, integrating genetically modified chondrocytes expressing transforming growth factor (TGF)‐β3 with ATDC5 cells, which demonstrated an augmented chondrogenesis effect. In this study, a delivery platform for the controlled release of stromal cell‐derived factor (SDF)‐1α was constructed based on our microcavitary hydrogel system. Subsequently, it was combined with the coculture system to explore the release patterns of SDF‐1α and TGF‐β3 and investigate their synergistic impact on chondrogenesis. The findings indicated that both SDF‐1α and TGF‐β3 could be continuously and efficiently released from the delivery system throughout the culture period. Moreover, the combined application of SDF‐1α and TGF‐β3 was able to enhance cell proliferative activity, as demonstrated by the Cell Counting Kit‐8 data. Synergistically, it led to superior chondrogenesis, as evidenced by real‐time PCR, Western blot analysis, and immunohistochemistry staining assays. This study offers the insight that this delivery system, integrating the coculture system and microcavitary hydrogels, holds substantial potential for cell recruitment and cartilage repair, and may be applicable in regenerative medicine for diverse tissues.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"644 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144513139","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}

{"title":"Magnetic‐Assisted Manipulation of Rare Blood Cells for Diagnosis: A Systematic Review","authors":"Poornima Ramesh Iyer, Xian Wu, Hyeon Choe, Linh Nguyen T. Tran, Karla Mercedes Paz González, Bahareh Rezaei, Shahriar Mostufa, Ebrahim Azizi, Ioannis H. Karampelas, Kai Wu, Jeffrey Chalmers, Jenifer Gomez‐Pastora","doi":"10.1002/bit.70010","DOIUrl":"https://doi.org/10.1002/bit.70010","url":null,"abstract":"The precise isolation and analysis of rare cells from blood are crucial for biomedical research and clinical diagnostics. This review examines recent advancements in magnetic‐based separation techniques, focusing on their efficiency in capturing rare cells such as circulating tumor cells (CTCs), circulating fetal cells, and diseased red blood cells (RBCs). These methods use magnetophoresis under external magnetic fields for highly specific isolation with minimal contamination, offering advantages over traditional techniques in speed, cost‐effectiveness, and robustness. Magnetic separation is categorized into label‐based methods, which use immunomagnetic nanoparticles (IMNs) to target specific cell markers, and label‐free methods, which exploit differences in magnetic susceptibility. Both approaches have achieved up to 99% efficiency in isolating diseased RBCs and CTCs. However, challenges remain in improving purity, scalability, and clinical applicability. A key limitation of label‐based methods is the need to detach cells from magnetic beads without compromising viability. Label‐free technologies, such as magnetic levitation, enable ligand‐free separation based on density and susceptibility. Future research should focus on optimizing paramagnetic media, integrating machine learning for enhanced accuracy, and developing high‐gradient magnetic fields (~1000 T/m) to improve efficiency. Advancements in IMNs with stronger magnetic properties will further enhance separation performance, driving clinical translation.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"7 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144500550","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}

{"title":"Identification of Lytic Enzymes With Activity Against Corynebacterium Species.","authors":"Monica A Mixon,Elena E Paskaleva,Sheila Luong,Marc P Douaisi,Mauricio Mora-Pale,Sandra P Sanchez-Rodriguez,Jonathan S Dordick","doi":"10.1002/bit.70009","DOIUrl":"https://doi.org/10.1002/bit.70009","url":null,"abstract":"Corynebacteriales are normal constituents of the human skin microbiota, involved in the metabolism of sweat and sebaceous glands secretions. The resultant compounds are a source of human body malodor. Some species, such as Corynebacterium jeikeium, are also opportunistic pathogens, and can cause systemic infection from colonization of medical devices such as intravascular catheters. Cell lytic enzymes have become effective alternatives to antibiotics and chemical oxidants in eliminating undesirable pathogens. As a baseline for antibacterial enzymatic activity we selected the broadly specific antibacterial enzyme lysozyme. While lysozyme had minimal activity on its own, the activity was significantly potentiated by the addition of a disruptor of the surface lipid layer of the bacterial cell wall, such as the lipase LysB (originating from a mycobacterial bacteriophage) or ethyleneglycol tetraacetic acid. To identify cell lytic enzymes with specific anti-Corynebacterial activity, we used a whole enzyme sequence as a probe to interrogate a custom-built in silico cDNA database of target Corynebacterial species and their bacteriophages. We also used a consensus enzymatically active domain as a probe. We evaluated the antibacterial activity of the thus discovered enzymes under a panel of application-relevant conditions, and determined that the newly identified bacterial autolysin NCBI WP_003861765.1 showed activity comparable to that of lysozyme, and the phage endolysin NCBI SLN03091 showed substantially higher activity than lysozyme and was significantly potentiated in the presence of LysB. As a result, we identified a nonchemical, nonantibiotic route to controlling Corynebacterial viability.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"55 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144488181","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}

{"title":"3D-Printed Sulfonated Polyetheretherketone Porous Scaffold Modified by Bioactive Glass for Bone Repair.","authors":"Xia Sheng,Zhenxu Wu,Xinyu Li,Qingming Ji,Xue Wang","doi":"10.1002/bit.70000","DOIUrl":"https://doi.org/10.1002/bit.70000","url":null,"abstract":"The bioinert characteristics of the polyetheretherketone (PEEK) matrix determine that the PEEK scaffolds cannot display bioactivity in bone regeneration. Scaffolds that combine the excellent mechanical strength of PEEK with the bioactivity of bioactive glass (BAG) are highly desirable for orthopedic applications. In this study, the porous structure of a PEEK scaffold was designed and fabricated via the fused deposition modeling (FDM) 3D printing method. The BAG was used as a coating material to cover the surface of sulfonated PEEK (SPEEK) via a novel chemical liquid deposition (CLD) method. The modified scaffold with a pore structure and rough surface promoted the mineralization on the scaffold. Additionally, the scaffold showed outstanding cytocompatibility and osteoactivity. Moreover, a bone defect repair study in rabbits revealed that the new bone tissues grew into the scaffold from the margin toward the center. The bone defect region was completely connected to the host bone end after 4 weeks of implantation. In summary, the SPEEK-BAG scaffold has promising potential for bone repair applications.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"20 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144478768","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}

{"title":"Biotechnological Advances in L-DOPA Biosynthesis and Production.","authors":"Hongmei Han,Yue Chen,Lingtian Wu,Yongsheng Wang,Yibo Zhu","doi":"10.1002/bit.70011","DOIUrl":"https://doi.org/10.1002/bit.70011","url":null,"abstract":"l-DOPA (3,4-dihydroxyphenyl-l-alanine) has been the primary medication for treating Parkinson's disease (PD), a degenerative brain disorder related to dopamine depletion, for the past six decades. As a result, biotechnological approaches utilizing metabolic engineering in microorganisms or enzymatic processes have been extensively explored as promising alternatives for l-DOPA production. These methods not only enhance conversion efficiency and enantioselectivity but are also cost-effective and environmentally sustainable. Metabolic engineering strategies have been employed to engineer Escherichia coli strains capable of accumulating l-DOPA from glucose by regulating carbon metabolism pathways. Additionally, microbial systems expressing tyrosinase, p-hydroxyphenylacetate 3-hydroxylase (PHAH), or tyrosine phenol-lyase (TPL) have been utilized for l-DOPA biosynthesis. In this review, we summarize current advancements in l-DOPA biosynthesis and biotechnological production strategies, providing a comparative analysis of their advantages and limitations. Moreover, we discuss the promise of biotech-driven l-DOPA production, emphasizing its industrial applications and large-scale production feasibility.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"178 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144370380","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}

{"title":"Improved Mixing Properties of Stirred Fermentation of an Aspergillus oryzae Hyphal Dispersion Mutant.","authors":"Shunya Susukida,Ken Miyazawa,Hikaru Ichikawa,Kiyoaki Muto,Akira Yoshimi,Toshitaka Kumagai,Yoshikazu Kato,Keietsu Abe","doi":"10.1002/bit.70004","DOIUrl":"https://doi.org/10.1002/bit.70004","url":null,"abstract":"The complexity of mechanical and biological processes in filamentous fungal fermentation remains a major obstacle to improving product yield. We previously demonstrated that the AGΔ-GAGΔ strain of Aspergillus oryzae, lacking both α-1,3-glucan (AG) and galactosaminogalactan (GAG), had improved hyphal dispersion, reduced culture viscosity, and increased recombinant protein production. Here, we applied computational fluid dynamics (CFD) and multi-omics analysis to characterize the AGΔ-GAGΔ strain during fermentation in a stirred-tank bioreactor. CFD simulations revealed large gas cavities behind the impeller blades and severe compartmentalization in both wild-type and AGΔ-GAGΔ cultures. However, shear stress distribution was broader and gas cavity formation was lower in the AGΔ-GAGΔ strain than in the wild type. The simulation results were consistent with measurements of volumetric oxygen mass transfer coefficients (KLa) and mixing times. Transcriptome analysis revealed upregulation of TCA-cycle genes in AGΔ-GAGΔ relative to the wild type. Analysis of intracellular and extracellular metabolites indicated distinct metabolic profiles associated with oxygen availability. Our findings highlight the critical role of hydrodynamics in fungal fermentation and demonstrate the potential of strain engineering for improving mixing characteristics.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"241 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144337500","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}

{"title":"Assessment of Anticancer Effects of Aloe vera on 3D Liver Tumor Spheroids in a Microfluidic Platform.","authors":"Atakan Tevlek,Gunes Kibar,Barbaros Cetin","doi":"10.1002/bit.29033","DOIUrl":"https://doi.org/10.1002/bit.29033","url":null,"abstract":"The search for effective anticancer therapies has increasingly focused on natural compounds like Aloe vera, renowned for its therapeutic properties. This study investigates the anticancer properties of Aloe vera on 3D liver tumor spheroids via a PDMS-based microfluidic device, providing a more physiologically realistic model compared to traditional 2D cultures. HepG2 cells were cultivated to generate 3D spheroids on-chip, thereafter subjected to different concentrations of Aloe vera and the chemotherapeutic drug Doxorubicin to evaluate cytotoxic effects. The microfluidic system, validated by COMSOL simulations, facilitated continuous perfusion and real-time assessment of cell viability over a duration of 10 days. The results indicated that Aloe vera markedly diminished cell viability by triggering apoptosis at concentrations over 12.5 mg/mL. IC50 values were determined at 72 h: 25 ± 0.10 mg/mL for Aloe vera and 5.47 ± 0.03 µg/mL for Doxorubicin in 2D cultures, but in 3D cultures, the IC50 values were 31.25 ± 0.14 mg/mL for Aloe vera and 8.33 ± 0.05 µg/mL for Doxorubicin. This study underscores the promise of Aloe vera as a natural anticancer agent and illustrates the efficacy of microfluidic platforms for enhanced drug screening and customized medicine applications.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"89 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144329069","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}

{"title":"The P134Q Sucrose Phosphorylase: Subtle Changes in the Catalytic Properties Benefit the Production of 2‐O‐α‐Glucosyl Glycerol","authors":"Alexander Sigg, Mario Klimacek, Martin Pfeiffer, Jorick Franceus, Tom Desmet, Bernd Nidetzky","doi":"10.1002/bit.70003","DOIUrl":"https://doi.org/10.1002/bit.70003","url":null,"abstract":"Protein engineering of <jats:italic>Bifidobacterium adolescentis</jats:italic> sucrose phosphorylase (<jats:italic>Ba</jats:italic>SucP) has previously identified the P134Q enzyme variant for site‐selective glycosylation at the 2‐OH of glycerol. Besides improvement in selectivity, the P134Q‐<jats:italic>Ba</jats:italic>SucP additionally involves enhanced affinity for glycerol as a biochemical property potentially important for the production of 2‐<jats:italic>O</jats:italic>‐α‐glucosyl glycerol (2GG), a commercialized skincare ingredient for cosmetic applications. Here, we performed a detailed kinetic model‐based evaluation of P134Q‐<jats:italic>Ba</jats:italic>SucP in initial‐rate and full reaction time course analyses to obtain a mechanistic interpretation and a comprehensive assessment of the process improvements achievable by the P134Q variant compared to the native enzyme. We show that P134Q‐<jats:italic>Ba</jats:italic>SucP involves ∼50‐fold lowered reactivity with phosphate compared to native enzyme. The effect likely arises from decreased conformational flexibility of the substrate binding pocket in the P134Q variant that may also serve to constrain the positioning of glycerol for glycosylation. Glycerol reactivity is decreased ∼1.3‐fold in P134Q‐<jats:italic>Ba</jats:italic>SucP; yet because the hydrolytic reactivity is lowered even more (threefold), the transfer efficiency to glycerol of the variant is enhanced ∼twofold compared to the native enzyme. Product inhibition by 2GG is decreased ∼threefold in P134Q‐<jats:italic>Ba</jats:italic>SucP. These properties of P134Q‐<jats:italic>Ba</jats:italic>SucP combine into major benefits for 2GG synthesis in terms of productivity and product yield. Model‐based window‐of‐operation analysis for 2GG production from sucrose and glycerol further reveals the significant potential for saving on the excess glycerol used in the process that results from replacing the wild‐type <jats:italic>Ba</jats:italic>SucP with the P134Q variant. Collectively, this study shows the important interplay of enzyme and reaction engineering in the optimization of glycoside production through biocatalytic transglycosylation.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"13 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144328598","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}