Biotechnology and Bioengineering最新文献

{"title":"Enrichment of Full AAV2 Using Multicolumn Countercurrent Solvent Gradient Purification (MCSGP)","authors":"Julia M. Müller,&nbsp;Tereza Müllerová,&nbsp;Daniela Tobler,&nbsp;Damian Hauri,&nbsp;Richard Plieninger,&nbsp;Yuki Higuchi,&nbsp;Ryosuke Takahashi,&nbsp;Sebastian Vogg,&nbsp;Thomas Müller-Späth,&nbsp;Thomas K. Villiger","doi":"10.1002/bit.29036","DOIUrl":"10.1002/bit.29036","url":null,"abstract":"<p>Adeno-associated viruses (AAVs) are non-pathogenic viruses that have become promising delivery vehicles in cell and gene therapy, with several AAV-based therapeutics receiving approval in recent years. However, a critical challenge is the separation of full and empty viral capsids, as empty capsids lack therapeutic DNA and may compromise product efficacy and safety. This study describes a proof-of-concept of a twin-column multicolumn countercurrent solvent gradient purification (MCSGP) process to enhance the ratio of full to empty capsids during AAV purification. Starting from a batch method with anion exchange chromatography, the process was adapted to continuous operation. The implementation of MCSGP, with six cycles, increased full capsid content in the final product pool from 30% to 68% (ddPCR) and 27% to 61% (cryo-TEM) respectively. Moreover, MCSGP was capable of overcoming the inherent yield-purity trade-off in the polishing step by enhancing full capsid recovery. Additionally, the MCSGP process improved productivity by up to 23% and reduced buffer consumption up to 27% compared to the batch method (ddPCR). The increased productivity and reduced buffer consumption offer both economic and environmental benefits. This study demonstrates the potential of MCSGP to meet the rising demand for high-quality AAV products in gene therapy.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2420-2432"},"PeriodicalIF":3.6,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.29036","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144219456","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":"Scalable Production of Recombinant Vesicular Stomatitis Virus Pseudoparticles Using HEK293 Suspension Cultures","authors":"Zhe Zhang,&nbsp;Elzbieta Wloga,&nbsp;Benjamin O. Fulton,&nbsp;Louis Coplan,&nbsp;Hanne Bak,&nbsp;Andrew D. Tustian","doi":"10.1002/bit.29042","DOIUrl":"10.1002/bit.29042","url":null,"abstract":"<div>\u0000 \u0000 <p>Recombinant vesicular stomatitis virus (rVSV) pseudoparticles displaying foreign glycoproteins are valuable for various applications, including vaccine vectors, oncolytic viruses, and research tools. Replication incompetent rVSV pseudoparticles, which do not encode their envelope proteins necessary for infection, offer increased versatility and rapid manufacturing. Traditionally, pseudotyping has been achieved using adherent host cells transiently expressing foreign glycoproteins, followed by infection of the host cells with a glycoprotein G (VSV-G) presenting virus that is deficient in glycoprotein genes. This adherent production method is challenging to scale up, which limits many applications. We developed a high-yield and scalable process using suspension adapted human embryonic kidney (HEK) 293F cells to produce replication incompetent rVSV. Using a multivariate approach, we optimized production duration to minimize the negative impact of residual transfection components and cellular waste products on viral infection and propagation. This eliminated the need for media exchange and enhanced process scalability. Key process parameters such as multiplicity of infection (MOI) and production duration were optimized to improve pseudoparticle productivity. The suspension process was scaled up to 2 L stirred tank bioreactors, yielding 7.4 × 10⁹ fluorescent forming units (FFU)/mL for VSV-G vector propagation and 2.4 × 10⁶ FFU/mL for pseudotyping with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein, achieving 8-fold and 50-fold higher productivity, respectively, than previous adherent processes. The pseudoparticles produced were fully neutralized by an anti-SARS-CoV-2 antibody, further validating the quality of the pseudoparticles from this suspension manufacturing process.</p>\u0000 </div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2353-2365"},"PeriodicalIF":3.6,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144211358","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":"Rational Design and Protein Engineering of {SH2 Domain–≫ Flexible Linker–≫ Self-Controlling Peptide} Fusion System With Phosphorylation-Regulated Molecular Switch Functionality","authors":"Peng Zhou,&nbsp;Yunyi Zhang,&nbsp;Kexin Li,&nbsp;Haiyang Ye,&nbsp;Li Mei,&nbsp;Shuyong Shang","doi":"10.1002/bit.29044","DOIUrl":"10.1002/bit.29044","url":null,"abstract":"<div>\u0000 \u0000 <p>Self-controlling peptides (SCPs) are described as a specific sub-class of our previously proposed self-binding peptides (SBPs), which work as a molecular switch to control the protein function conversion between two or more biological states. In this study, a variety of artificial protein systems containing SCP with molecular switch functionality were rationally designed and systematically engineered by fusing several (weak and strong) phosphopeptide binders to the C-terminal tail of human Src SH2 domain via a flexible linker (FL), termed {SH2–&gt; FL–&gt; SCP} fusion protein systems. It is revealed that the intramolecular interaction between SH2 and SCP in the systems shares a similar binding behavior with the intermolecular interaction between SH2 and free phosphopeptide in structural and energetic points of view. In addition, FL is responsible for the binding dynamics of SCP to SH2, which restricts the SCP in a local region nearby SH2 binding site, thus equivalently enhancing the micro-concentration of SCP around the site, effectively increasing the collision frequency of SCP with the site, and consequently improving the apparent affinity of SCP to SH2. We carefully selected phosphopeptide binders and systematically optimized the sequence length and amino acid composition of FL, consequently resulting in a satisfactory {SH2–&gt; poly(G)₁₂–&gt; SCP(SIPM2-K_-2)} fusion protein system that was demonstrated to possess molecular switch functionality, in which the SCP binding and unbinding events were reversibly triggered by externally controlling its Tyr0-phosphorylation and pTyr0-dephosphorylation, respectively.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2511-2521"},"PeriodicalIF":3.6,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144219458","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":"A Scalable Deep Learning Approach for Real-Time Multivariate Monitoring of Biopharmaceutical Processes With No Prior Product-Specific History","authors":"Nima Sammaknejad,&nbsp;Jessica Lee,&nbsp;Jan Michael Austria,&nbsp;Nadia Duenas,&nbsp;Leila Heiba,&nbsp;Govi Sridharan,&nbsp;Jeff Davis,&nbsp;Cenk Undey","doi":"10.1002/bit.29039","DOIUrl":"10.1002/bit.29039","url":null,"abstract":"<div>\u0000 \u0000 <p>Real-time multivariate statistical process monitoring (RT-MSPM) is essential to monitor health of bio-pharmaceutical processes and detect anomalies and faults early in the process. RT-MSPM methods are commonly used to monitor cell culture process operations in biologics drug substance manufacturing. Batch evolution models (BEMs) are among common RT-MSPM methods. As an alternative to BEMs, it is possible to develop multiple models to monitor different phases of a batch process. If certain statistical properties are satisfied, a multistage algorithm can be leveraged to detect steady state operation of a batch and process the corresponding time-series in a manner to leverage data from other product recipes to monitor a new product with no prior history. This is specifically useful in modern biopharmaceutical manufacturing facilities, which frequently switch from producing one medicine to another. In this article, a novel real-time deep learning framework to monitor the health of biopharmaceutical processes with no prior product-specific history is proposed. Autoencoders (AEs), in conjunction with a multistage real-time data processing algorithm, are leveraged to detect, prevent and identify the root causes of potential anomalies and faults in cell culture manufacturing processes to produce monoclonal antibodies with no prior history. A novel algorithm for real-time root cause identification of anomalies is developed to generate real-time contribution charts for AEs. The performance of the new fault detection and isolation strategy is compared with conventional methods. Given the nonlinear architecture of AEs in comparison to conventional linear methods, AEs consistently provide more robust and stronger evidence for anomalous patterns using a combination of information in residuals and latent space. The proposed framework is successfully tested within a scalable software product for real-time monitoring of manufacturing cell culture bioreactors.</p>\u0000 </div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2333-2352"},"PeriodicalIF":3.6,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144202197","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":"Integration of Single Pass Tangential Flow Filtration and High Performance Countercurrent Membrane Purification for Intensification of Monoclonal Antibody Processing","authors":"Mirko Minervini,&nbsp;Aylin Mohammadzadehmarandi,&nbsp;Ali Behboudi,&nbsp;Sheldon F. Oppenheim,&nbsp;Andrew L. Zydney","doi":"10.1002/bit.29038","DOIUrl":"10.1002/bit.29038","url":null,"abstract":"<p>There is increasing interest in the development of intensified processes that can provide significant reductions in manufacturing costs for monoclonal antibody (mAb) products. This study examines the performance of an integrated membrane process that combines single-pass tangential flow filtration (SPTFF) with high-performance countercurrent membrane purification (HPCMP) for the preconditioning of clarified culture fluid (CCF). The SPTFF step was operated with a single-pass concentration factor of 20x for 24 h filtration, with no evidence of membrane fouling; the transmembrane pressure remained &lt; 30 kPa throughout the process. HPCMP was then performed on the pre-concentrated feed, resulting in a 60 ± 8% reduction in host cell proteins (HCPs) and more than a 30-fold removal of DNA while maintaining 94 ± 3% mAb step yield. The HPCMP process operated continuously for 36 h with a slight decline in HCP removal in the latter stages of operation due to membrane fouling. The buffer requirement for the integrated process was only 76 L/kg mAb. These results demonstrate the potential of using an integrated SPTFF-HPCMP process for preconditioning CCF as part of a highly intensified mAb manufacturing process with much lower cost of goods.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2410-2419"},"PeriodicalIF":3.6,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.29038","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144198247","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":"Dynamic Regulation of Efflux Pump Protein AcrA by Quorum Sensing System to Improve Free Fatty Acids Production in Escherichia coli","authors":"Lixia Fang,&nbsp;Peishi Wen,&nbsp;Jiaqi Zhang,&nbsp;Yingxiu Cao","doi":"10.1002/bit.29041","DOIUrl":"10.1002/bit.29041","url":null,"abstract":"<div>\u0000 \u0000 <p>Microbial production of free fatty acids (FFAs) is an eco-friendly and promising approach. However, FFAs accumulation within microbial cells imposes stress and toxicity, impairing biosynthetic performance. These challenges can be alleviated by exporting FFAs through the efflux system AcrAB-TolC in <i>Escherichia coli</i>, but the expression of these membrane channels needs to be fine-tuned. In this study, we employed a quorum sensing (QS) system to dynamically regulate the expression of AcrAB-TolC, thereby enhancing FFAs efflux and production. Two regulatory patterns of gene expression were designed to respond to increasing cell density: EsaI/R-P_<i>esaS</i> drives an initial increase followed by a decrease pattern, and EsaI/R-P_<i>esaR</i> enables a slow linear increase pattern. The L19IR-P_R-<i>acrA</i> strain, in which <i>acrA</i> expression is auto-induced by P_<i>esaR</i>, exhibited a 142% increase in extracellular FFAs and an 11% increase in total FFAs titer compared to the L19IR strain, which lacks <i>acrA</i> regulation. Auto-induced dynamic regulation of <i>acrA</i> demonstrates significant improvements in both FFAs efflux and overall production, while maintaining cell growth and membrane stability. Our results highlight the potential of QS-mediated dynamic regulation of efflux pumps to enhance the synthesis of bioproducts toxic to microorganisms.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2499-2510"},"PeriodicalIF":3.6,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144179649","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":"Techno-Economic Analysis of Membrane-Based Purification Platforms for AAV Vector Production","authors":"Juan J. Romero,&nbsp;Eleanor W. Jenkins,&nbsp;Jacob I. Monroe,&nbsp;S. Ranil Wickramasinghe,&nbsp;Xianghong Qian,&nbsp;Dibakar Bhattacharyya,&nbsp;Scott M. Husson","doi":"10.1002/bit.29034","DOIUrl":"10.1002/bit.29034","url":null,"abstract":"<p>Technologies for large-scale manufacturing of viral vectors for gene therapies, such as tangential flow filtration and membrane chromatography, are under development. In these early stages of process development, techno-economic analyses are useful for identifying membrane properties yielding the greatest impact on process performance. In this study, we adapted a techno-economic framework used for monoclonal antibody capture for adeno-associated viral vector purification. We added mechanistic models to simulate flux decline during harvesting and separating full and empty capsids during polishing. Graphical user interfaces were added to help users explore the design search space. We selected a base process and manipulated selected variables to see their impact on large-scale manufacturing performance. These sensitivity analyses revealed that, under the selected process conditions, increasing module capacity reduces cost of goods more effectively than increasing operational flux in tangential flow membrane filtration modules for virus harvesting. Membrane chromatography columns with relatively low dynamic binding capacity (DBC) and short residence time (RT) offered similar or better economic performance than those with high DBC and long RT. Additionally, the difference in equilibrium solid-phase concentration between full and empty capsids as a function of salt concentration significantly affects purity.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2400-2409"},"PeriodicalIF":3.6,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.29034","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144180296","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":"N- to C-Glycoside Rearrangement of Uridine 5′-Phosphate in Two Enzymatic Steps for the Production of Pseudouridine 5′-Phosphate","authors":"Martin Pfeiffer,&nbsp;Franziska Guld,&nbsp;Bernd Nidetzky","doi":"10.1002/bit.29037","DOIUrl":"10.1002/bit.29037","url":null,"abstract":"<p>Pseudouridine (<b>Ψ</b>) is an essential building block of synthetic RNA for medical applications, so methods for its efficient production receive increased interest. Reverse reaction of the <b>Ψ</b>-5′-phosphate (<b>ΨMP</b>) <i>C</i>-glycosidase, that is, <span>d</span>-ribose 5-phosphate (<b>Rib5P</b>) + uracil (Ura) → <b>ΨMP</b> + H₂O, allows for the installment of the core β-<i>C</i>-riboside structure of <b>Ψ</b> in a completely selective and efficiently equilibrium-driven single-step transformation. However, providing the <b>Rib5P</b> substrate is challenging for process development and optimum solutions can vary depending on the specific production tasks considered. Here, we exploited the less known activity of purine/pyrimidine nucleotide 5′-phosphate nucleosidase (PpnN; EC 3.2.2.10) to cleave uridine 5′-phosphate (<b>UMP</b>), a relatively expedient starting material for <b>ΨMP</b> synthesis, under release of <b>Rib5P</b> and <b>Ura</b>. Using linear cascade transformation in two enzymatic steps performed in one pot, we demonstrate rearrangement of <b>UMP</b> into <b>ΨMP</b> (yield: ≥ 95%) and thereby obtain the <i>C</i>-riboside product at the solubility limit (∼1.5 mol/L) in a productivity of 2.9 × 10²g/L/h. We show that a previously reported R341A-Y347A double variant of <i>Escherichia coli</i> PpnN (RY) exhibited ∼5-fold higher specific activity toward UMP, and was ∼12-fold less sensitive to <b>Rib5P</b> inhibition, than the wild-type enzyme. Under conditions of <i>C</i>-glycosidase applied in twofold excess over PpnN to minimize the effect of <b>Rib5P</b> inhibition, cascade reaction with RY compared to wild-type PpnN still gave ∼5-fold enhanced productivity. In summary, we present a new synthetic route to <b>ΨMP</b> via <i>N</i>- to <i>C</i>-glycoside rearrangement of <b>UMP</b>. Compared to earlier approaches of cascade biocatalysis for <b>ΨMP</b> production from uridine or <b>UMP</b>, this new route is streamlined due to the direct release of <b>Rib5P</b> from the <b>UMP</b> substrate catalyzed by PpnN.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2456-2464"},"PeriodicalIF":3.6,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.29037","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144156532","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":"Dynamic Compression Improves Chondrogenesis in the Tissue Engineered Model of Cartilage","authors":"Marc V. Farcasanu,&nbsp;Thais de las Heras Ruiz,&nbsp;Francesca M. Johnson de Sousa Brito,&nbsp;Jamie Soul,&nbsp;Jonathan Coxhead,&nbsp;Matthew J. German,&nbsp;David A. Young,&nbsp;Ana M. Ferreira-Duarte,&nbsp;Katarzyna A. Piróg","doi":"10.1002/bit.29026","DOIUrl":"10.1002/bit.29026","url":null,"abstract":"<p>Hyaline cartilage is a dense avascular tissue with low regenerative potential, present at the ends of the diarthrodial joints and in the cartilage growth plate. Skeletal diseases often result from extracellular changes in this tissue; however, studies of these are hindered by the tissue complexity, the difficulty in obtaining human material, and the cost of generating animal models. Recent developments in tissue engineering are opening possibilities to develop mechanoresponsive zonally stratified models of cartilage in vitro. In this study, we optimized a 3D model of cartilage using chondroprogenitor cells cultured for 21 days in 2% agarose hydrogel constructs with daily dynamic compression. Our hydrogel constructs developed pericellular matrices with nanostiffness comparable with native murine tissue and showed increased production of extracellular matrix components and expression of chondrogenic and differentiation markers. Daily dynamic compression resulted in progressive increase in mechanoresponsive gene expression and promoted a juvenile cartilage phenotype, decreasing expression of dedifferentiation and cartilage degradation markers. Our study highlights the potential of hydrogel-enhanced chondrogenesis and proposes an adaptable and scalable in vitro model to study mechanoresponses, intracellular signals, and pericellular matrix involvement in cartilage development and disease.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2574-2591"},"PeriodicalIF":3.6,"publicationDate":"2025-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.29026","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144136823","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":"Post-Treatment of CO₂ Emissions With Microalgae: Magnetic Field-Induced Improvements in an AirLift Photoreactor","authors":"Luis Torres,&nbsp;José Octavio Saucedo-Lucero,&nbsp;Aitor Aizpuru,&nbsp;Sonia Arriaga","doi":"10.1002/bit.29032","DOIUrl":"10.1002/bit.29032","url":null,"abstract":"<div>\u0000 \u0000 <p>Atmospheric pollution from volatile organic compounds (VOCs) and rising global temperatures due to greenhouse gases (GHGs) emissions, such as carbon dioxide (CO₂) pose significant threats to air quality and public health. Coupled biological systems can mitigate VOC emissions, generating CO₂, which is then assimilated by microalgae. Static magnetic field (SMF) stimulation has been shown to enhance microalgal growth and CO₂ fixation. This study evaluated the impact of SMF on CO₂ fixation in an airlift photoreactor (ARL) following VOCs treatment in a semi-continuous stirred tank reactor (S-CSTR) processing toluene vapors. The ARL was exposed to SMF at 45 mT for 6, 4, and 2 h d⁻¹. Results demonstrated a 96% increase in CO₂ capture after 4 h of exposure, while removing 45% of the remanent toluene. The highest biomass increase (12%) occurred after 6 h of exposure, whereas total chlorophyll content peaked at 18.4 mg L⁻¹ under 4 h of SMF, compared with 6.8 mg L⁻¹ in the control. Therefore, 4 h exposure at 45 mT was identified as the optimal condition, balancing VOCs reduction, CO₂ mitigation, and high pigment production. Microalgal cultures under SMF present a promising and versatile approach for air pollution control and carbon valorization, offering potential economic benefits through biomass applications and supporting circular economy initiatives.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 9","pages":"2319-2332"},"PeriodicalIF":3.6,"publicationDate":"2025-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144136696","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}