Biotechnology and Bioengineering最新文献

{"title":"Nanocarrier-Based Gene Delivery Systems: Mechanisms, Clinical Translation, and Future Perspectives","authors":"Rahat Fatima Naqvi,&nbsp;Muhammad Ali,&nbsp;Sadia Ahmed Zuberi,&nbsp;Nimra Aamir,&nbsp;Samra Rafique","doi":"10.1002/bit.70289","DOIUrl":"10.1002/bit.70289","url":null,"abstract":"<div>\u0000 \u0000 <p>Gene therapy holds revolutionary potential for managing genetic disorders, cancers and infectious illnesses. However, one of the biggest challenges is delivering DNA or RNA into targeted cells and in the safe and effective way. In this review, nano carrier-based approaches for gene delivery are critically examined, focusing on both viral and non-viral systems. The advancement of CRISPR-Cas genome editing, machine learning-assisted nanocarrier optimization, and biologically inspired delivery systems is being quickly pushed forward in this area. In this review, a comparative analysis of gene delivery systems is being provided, and the key challenges to clinical translation are being pointed out. In addition, expert opinions on future research directions are being offered, with a heavy focus on the development of multifunctional, precisely targeted, and easily scalable delivery systems that can be integrated with next-generation therapeutic technologies.</p>\u0000 </div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2553-2568"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148342272","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":"Bacterial Laccase: Recent Advances in Production, Engineering Strategies, and Diagnostic Applications","authors":"Fina Wulandari Mardiyono,&nbsp;Bunga Septiyadi Aulia Putri,&nbsp;Asrul Muhammad Fuad,&nbsp;Neng Herawati,&nbsp; Desriani,&nbsp;Nuruliawaty Utami,&nbsp;Shabarni Gaffar,&nbsp;Dessy Natalia,&nbsp; Widoretno,&nbsp;Sri Harjati Suhardi,&nbsp;Ernawati Arifin Giri-Rachman,&nbsp;Dani Permana","doi":"10.1002/bit.70272","DOIUrl":"10.1002/bit.70272","url":null,"abstract":"<div>\u0000 \u0000 <p>Bacterial laccases are multicopper oxidase enzymes that are able to catalyze the oxidation reaction of phenolic and non-phenolic substrates followed by the reduction reaction of molecular oxygen to water as the only byproduct. It exhibits high stability under harsh conditions and can be produced recombinantly in various microbial hosts. Herein, we review recent advances in bacterial laccase research over the past decade, which focus on production technologies, protein engineering strategies, and diagnostic applications. We also discuss how synthetic biology has overcome historical limitations in heterologous expression and enabled industrial-scale production in hosts such as <i>Escherichia coli</i> and <i>Bacillus subtilis</i>. In addition, we analyzed how directed evolution, rational design, post-translational modifications, and computational modeling have enhanced the catalytic properties of laccases. In the final chapter, we evaluate the use of bacterial laccases as peroxidase alternatives for detecting clinical analytes like dopamine, uric acid, and glucose biosensing and point-of-care diagnostics. This review integrated more than 120 peer-reviewed papers and provides a comprehensive assessment of how bacterial laccases are transitioning from laboratory curiosities to practical biocatalysts in environmental monitoring, industrial processing, and medical diagnostics. We conclude the review by identifying remaining challenges and future research directions for realizing the full potential of these versatile enzymes.</p>\u0000 </div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2433-2452"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148270415","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":"Cardiovascular Organoids With Adjustable Endothelial Composition via SOX17-Engineered hPSCs","authors":"Po-Yu Liang,&nbsp;Gyuhyung Jin,&nbsp;Nathan R. Petrucci,&nbsp;Xiaojun Lance Lian,&nbsp;Xiaoping Bao","doi":"10.1002/bit.70275","DOIUrl":"10.1002/bit.70275","url":null,"abstract":"<p>Organoids are considered a novel modeling platform for studying human biology and advancing health research. With the ability to demonstrate complex 3D structure and multicellular interactions, organoids have advanced studies in all major organs as a reliable model. In this study, we generated an advanced cardiovascular organoid by using a genome-edited human pluripotent stem cell line with inducible <i>SOX17</i> expression, enabling controlled endothelial specification, adjustable cell-type composition, and human heart-like morphology. Our organoids recapitulated the cardiotoxic phenotypes of FDA-approved chemotherapeutic doxorubicin, manifesting as decreased cell viability and diminished contractile activity. Cryoinjury-induced myocardial infarction in our organoids led to reduced beating, viability, and α-actinin expression, along with increased fibroblast formation, which were mitigated by Captopril. Lastly, isoproterenol treatment increased peak Ca²⁺ transient amplitude and shortened APD₅₀ in our organoids, consistent with previously reported β-adrenergic responses. In summary, we established a protocol for generating in vitro 3D cardiovascular organoids with controllable cellular composition and heart-like structures, providing a robust and easy-to-produce platform for future studies of human heart disease.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2655-2667"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70275","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148288242","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic Flux Analysis Reveals Cell Line-Specific Rewiring in CHO Cells Following TCA Cycle Intermediate Feeding for Bioprocess pH Control","authors":"Sarah A. Sacco,&nbsp;Kevin J. Ruiz-Márquez,&nbsp;Rachel Moen,&nbsp;Irina Trenary,&nbsp;Xiaolin Zhang,&nbsp;Xiaowen Wang,&nbsp;Malik Padellan,&nbsp;Henry Lin,&nbsp;Jamey D. Young","doi":"10.1002/bit.70287","DOIUrl":"10.1002/bit.70287","url":null,"abstract":"<p>Chinese hamster ovary (CHO) cells are the leading host for recombinant therapeutic protein production in the biopharma industry. In this study, we investigated how feeding acidic forms of tricarboxylic acid (TCA) cycle intermediates—malic acid, succinic acid, and α-ketoglutaric acid—affects cell culture performance and metabolism in two industrial IgG-producing CHO cell lines. These intermediates were used as pH control agents to replace conventional CO₂ sparging, enabling simultaneous modulation of the bioreactor environment and cell metabolism. Carbon-13 metabolic flux analysis (¹³C-MFA) revealed substantial rewiring of central carbon and nitrogen metabolism under all fed conditions, with distinct responses between cell lines. Intermediate-fed cultures exhibited enhanced TCA cycle fluxes, reduced glucose dependency, decreased lactate accumulation, and altered routing of pyruvate. Notably, α-ketoglutaric (α-KG) acid feeding triggered divergent nitrogen assimilation phenotypes: one cell line enhanced glutamine biosynthesis and ammonium clearance, while the other accumulated glutamate with minimal glutamine production. These metabolic adaptations were accompanied by shifts in redox balance and delayed but measurable increases in cell-specific IgG productivity. Our findings highlight the compound- and cell line-specific nature of metabolic responses to TCA cycle intermediate feeding and support its use for pH control and bioprocess optimization.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2713-2728"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70287","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148342251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Spray-Dried Engineered Escherichia coli Whole-Cell Biocatalysts Enable Cell Recycling and In Vivo α-Ketoglutarate and NADPH Regeneration for Efficient Codeine Manufacture","authors":"Ali Jahanian,&nbsp;Carlos Horacio Luna-Flores,&nbsp;Xu Li,&nbsp;Sally L. Gras,&nbsp;Robert E. Speight","doi":"10.1002/bit.70270","DOIUrl":"10.1002/bit.70270","url":null,"abstract":"<p>A recombinant whole-cell biocatalytic approach provides several advantages over isolated enzymes, such as improved enzyme stability, in vivo cofactor regeneration, and the facilitation of cascade reactions within a single cell. Whole-cell biocatalysis has gained attention in medicinal opioid production, due to its technical and economic advantages. In this study, recombinant <i>Escherichia coli</i> producing thebaine 6-<i>O</i>-demethylase (T6ODM), neopinone isomerase (NISO), and codeinone reductase (COR) were used to convert thebaine into codeine. Spray-drying was developed to optimize enzymatic activity in dried biocatalytic cells, achieving over 85% enzyme activity retention. These formulated cells were evaluated over multiple recycled reaction cycles that relied on in vivo cofactor regeneration. The native enzyme isocitrate dehydrogenase (IDH) regenerated both α-ketoglutarate and NADPH, critical for T6ODM and COR activity, respectively. The biocatalysts demonstrated high durability across 10 reaction cycles, with IDH and COR maintaining consistent activity. However, T6ODM exhibited a 40% reduction in activity, aligning with its lower thermal stability. The two-step bioconversion process yielded an average 68% molar yield of codeine after 10 cycles, with full thebaine consumption, demonstrating efficient cofactor and co-substrate recycling without further cellular engineering. This study highlights the potential of spray-dried <i>E. coli</i> cells as a sustainable and scalable approach for codeine biomanufacturing from thebaine. These results provide a strong foundation for further industrial application, offering both economic and technical benefits for large-scale opioid production.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2629-2642"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70270","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148216445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and Integrated Application of the Multi-Attribute Method (MAM) in Quality Control of Biotechnological Drugs","authors":"Yuan Zhu,&nbsp;Doudou Lou,&nbsp;Chen Yang,&nbsp;Ran Ding,&nbsp;Min Song,&nbsp;Rong Wang,&nbsp;Yihong Lu,&nbsp;Qingfeng Fan","doi":"10.1002/bit.70274","DOIUrl":"10.1002/bit.70274","url":null,"abstract":"<div>\u0000 \u0000 <p>The multi-attribute method (MAM) is an integrated peptide mapping strategy based on liquid chromatography-mass spectrometry (LC-MS) technology. It enables precise quantification and dynamic tracking of multiple site-specific modifications in a single analysis, significantly enhancing the efficiency and depth of biopharmaceutical quality control. Notably, its integrated application across process development, process monitoring, and product release has driven a paradigm shift from a “single-attribute, single-method” approach to a “multi-attribute, integrated-method” approach in quality control. This review systematically summarizes the technical principles, optimization strategies, and application progress of MAM by integrating recent research cases of complex therapeutic proteins (e.g., monoclonal antibodies and Fc fusion proteins), with a focus on specific strategies and practical paths of MAM in workflow automation, new peak detection (NPD) optimization, intact multi-attribute method (iMAM), and the integration of complementary technologies. The objective is to provide a valuable reference for the standardization and industrial application of MAM in biotechnological drug quality control. Although MAM is expected to become a core analytical tool for biopharmaceutical quality control, its widespread industrial application remains constrained by key challenges, including insufficient method robustness, incomplete standardization, and variable regulatory acceptance. Notably, a significant stride in regulatory acceptance has been made with the recent implementation of the United States Pharmacopeia (USP) General Chapter &lt; 1060 &gt; , which establishes the first official framework for MAM. Beyond this regulatory milestone, future efforts should focus on advancing automated platform development, creating intelligent data algorithms, and strengthening cross-disciplinary collaboration to further promote the systematic integration and standardized application of MAM throughout the full lifecycle quality management of biotechnological drugs.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2536-2552"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148342273","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":"Microbial Hydrolysates as Amino Acid Source in Cell Culture Media for Cellular Agriculture","authors":"Jelle C. Hoek,&nbsp;Jean-Marc Daran,&nbsp;Marieke E. Klijn,&nbsp;Marcel A. Vieira-Lara","doi":"10.1002/bit.70290","DOIUrl":"10.1002/bit.70290","url":null,"abstract":"<p>The high cost of cultivation media remains one of the barriers to the commercialization of cultivated meat. Current serum-free media reduce reliance on fetal bovine serum but only achieve modest cost savings. Especially at larger scales (&gt; 20 m³), pharmaceutical-grade amino acids and recombinant proteins remain major cost drivers. Techno-economic analyses suggest that replacing purified amino acids is essential to achieve price parity with conventional meat. Microbial hydrolysates, also referred to as extracts, offer a promising alternative, as these can supply complex mixtures of amino acids and peptides at lower cost and with greater scalability than chemically defined formulations. In particular, yeast and bacterial extracts combine high protein (and therefore high amino acid) content, rapid growth, and established industrial-scale production. Comparative analyses indicate that microbial amino acid profiles broadly overlap with those of animal cells and traditional meat, but this similarity does not translate into meeting cellular amino acid demand. Consumption data indicate that key amino acids, such as glutamine, cysteine, serine and arginine, would be supplied insufficiently by microbial extracts. This mismatch highlights the need for engineering of microbial biomass and optimization of extraction methods. Extraction methods such as autolysis, enzymatic lysis, or physical disruption strongly influence nutrient release and composition, underscoring the need for standardizing microbial extract preparation. In addition, challenges remain in ensuring consistency, safety, and bioavailability of nutrients, as microbial extracts also contain nucleic acids and potential toxins. This review summarizes current knowledge on microbial hydrolysates for cultivated meat media, including biomass composition, extraction methods, and analytical tools to assess quality and performance. We identify key knowledge gaps, particularly in quantitative amino acid consumption data for relevant cell lines and performance testing in scalable cultures. Addressing these gaps could enable cost-effective media development and accelerate research toward sustainable, commercially viable cultivated meat.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2520-2535"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70290","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148372696","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Highly Efficient Enzymatic Catalytic Transformation for the Synthesis of Rare Ginsenoside Compound K (CK) and Its Bioactivity Study","authors":"Hao Tang,&nbsp;Shihua Han,&nbsp;Zidan Li,&nbsp;Yiwen Jiang,&nbsp;Meimei Fu,&nbsp;Zhuoyi Huang,&nbsp;Lei Jia,&nbsp;Fengrui Song,&nbsp;Jianguo Huang,&nbsp;Gloria B. Kim,&nbsp;Jinshan Guo,&nbsp;Zhihui Lu","doi":"10.1002/bit.70277","DOIUrl":"10.1002/bit.70277","url":null,"abstract":"<div>\u0000 \u0000 <p>As a principal metabolite of ginsenosides, compound K (CK) exhibits remarkable antioxidant, anti-inflammatory, antidiabetic, and anticancer bioactivities. However, CK is naturally scarce in ginseng and its industrial application is limited by the lack of efficient deglycosylating enzymes for large-scale CK bioproduction. Here, we report a synthetic biology-driven approach for high-yield CK production <i>via</i> recombinant β-glycosidase-mediated bioconversion of the polar ginsenoside Rb1. The β‑glycosidase-encoding gene was introduced into <i>Escherichia coli</i> BL21 (DE3), and the resulting engineered strain produced 2.1 g/L β‑glycosidase in shake flasks and 10.1 g/L in a 5-L fermentor. The crude β-glycosidase exhibited excellent enzymatic activity; at a concentration of 1.5 g/L, it converted 95% of the Rb1 substrate (initial concentration 10 g/L) to CK within 4 h. <i>In vitro</i> studies demonstrated CK's high cytocompatibility and potent anti-aging effects, including suppression of reactive oxygen species (<i>via</i> upregulation of superoxide dismutase, catalase, and glutathione peroxidase), enhanced collagen I, Ⅲ and elastin synthesis, and matrix metalloproteinase-1 inhibition. These findings highlight CK as a promising candidate for anti-wrinkle formulations, supported by the feasibility of scalable bioproduction.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2668-2676"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148522374","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":"Biotechnology and Bioengineering: Volume 123, Number 10, October 2026","authors":"","doi":"10.1002/bit.70388","DOIUrl":"https://doi.org/10.1002/bit.70388","url":null,"abstract":"","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2429-2432"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.70388","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148965561","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A CRISPR-Cas9 Toolkit Enabling Tunable Integration and Transient Homologous Recombination Enhancement in Yarrowia lipolytica","authors":"Tang-Lei Zhang,&nbsp;Ming-Kai Chen,&nbsp;Yue-Min Li,&nbsp;Hong-Wei Yu,&nbsp;Li-Dan Ye","doi":"10.1002/bit.70276","DOIUrl":"10.1002/bit.70276","url":null,"abstract":"<div>\u0000 \u0000 <p>Although the oleaginous yeast <i>Yarrowia lipolytica</i> is a promising microbial cell factory, its application remains constrained by inefficient homology-directed repair (HDR) and a lack of precise genomic integration tools. To address these limitations, we developed a comprehensive genetic toolkit featuring three synergistic advancements. First, we systematically identified 55 neutral integration sites with tunable expression profiles, enabling stable, position-independent gene integration with predictable transcriptional output across a 12.88-fold dynamic range. Second, we established a dual-readout high-throughput screening platform combining colony morphology analysis with hrGFP fluorescence. This approach accurately measures locus-specific homologous recombination (HR) efficiency while eliminating false positives by dominant non-homologous end joining (NHEJ). Third, we engineered a transient HR enhancement system by fusing the Sae2 exonuclease to Cas9 via a flexible (GGGGS)₃ linker. This fusion significantly boosts HR efficiency and surpasses the cleavage activity of unmodified Cas9 without introducing permanent genomic modifications or compromising cellular fitness. Finally, HR efficiency for single-gene integration was increased from 46.5% to 77.5% while the dual-locus editing efficiency reached 64.1% when using 500-bp homology arms, and the engineered strains demonstrated improved genetic stability compared to those with constitutive HR enhancement.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"123 10","pages":"2643-2654"},"PeriodicalIF":3.9,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148288239","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}