Biotechnology Journal最新文献

{"title":"Cell Seeding Strategy Influences Metabolism and Differentiation Potency of Human Induced Pluripotent Stem Cells Into Pancreatic Progenitors","authors":"Hui Huang,&nbsp;Kaiming Ye,&nbsp;Sha Jin","doi":"10.1002/biot.70022","DOIUrl":"https://doi.org/10.1002/biot.70022","url":null,"abstract":"<p>Human induced pluripotent stem cells (iPSCs) are an invaluable endless cell source for generating various therapeutic cells and tissues. However, their differentiation into specific cell lineages, such as definitive endoderm (DE) and pancreatic progenitor (PP), often suffers from poor reproducibility, due partially to their pluripotency. In this work, we investigated the impact of iPSC confluency during cell self-renewal and seeding density on cell metabolic activity, glycolysis to oxidative phosphorylation shift, and differentiation potential toward DE and PP lineages. Our findings demonstrated that cell seeding strategy influences cellular metabolic activity and the robustness of iPSC differentiation. iPSCs maintained at higher seeding density exhibited lower initial oxygen consumption rate (OCR) and metabolic activity. There is an optimal seeding density to ensure sufficient oxygen consumption during differentiation and to yield high expression of SOX17 in the DE lineage and high PDX1/NKX6.1 dual-positive cells in PPs. Interestingly, we found that cell confluency at the time of harvest has less impact on the efficacy of pancreatic lineage formation or metabolic activity. This study sheds light on the interplay between metabolic activity and iPSC lineage specification, offering new insights into the robustness of iPSC self-renewal and differentiation for creating human tissues.</p>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70022","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875656","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":"Impact of Eco-Friendly Surfactant Structure and Class on Enveloped Virus Inactivation","authors":"Vaishali Sharma,&nbsp;Lynn Manchester,&nbsp;Melissa Holstein,&nbsp;Xuankuo Xu,&nbsp;Sanchayita Ghose,&nbsp;Caryn L. Heldt","doi":"10.1002/biot.70023","DOIUrl":"https://doi.org/10.1002/biot.70023","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Sustainable and effective strategies for virus inactivation are crucial for ensuring the safety and quality of biological products. The European Union's (EU) 2021 ban on Triton X-100 for viral inactivation in biomanufacturing has pushed the field to find sustainable alternatives with equal effectiveness. We aim to increase the sustainability of biopharmaceutical production by ensuring the effectiveness of eco-friendly surfactant-mediated virus inactivation by comparing the antiviral efficacy of Triton X-100 to glucosides and amine oxides.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Surfactants were evaluated for antiviral efficacy against herpes viruses, SuHV and HSV, and the retrovirus XMuLV. The surfactants demonstrated equivalent or superior inactivation efficacy compared to Triton X-100. Herpes viruses were inactivated similarly with all surfactants. For XMuLV, surfactants with longer alkyl chains achieved maximum log reduction values (LRV) at 1x CMC, outperforming Triton X-100, which required 2x CMC for comparable efficacy. Surfactants with bulky headgroups, such as LAPAO, showed lower efficacy against XMuLV. At a salt concentration of 2 M ionic strength, the antiviral efficacy of Triton X-100 and TDAO decreased for the herpes viruses. Variability in inactivation was observed among the surfactants at 0.5x CMC, indicating that surfactant characteristics influence their antiviral performance below CMC.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>Adding salt enhanced the antiviral efficacy of surfactants by lowering their CMC while maintaining consistent virus inactivation. Among the surfactants tested, the glucoside with a longer tail, n-nonyl-β-D-glucoside (NG), emerged as the most robust and could function as an eco-friendly surfactant for virus inactivation in bioprocessing. For NG, virus inactivation was independent of all variables tested.</p>\u0000 </section>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875593","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrating Biofilm-Based Fermentation With Continuous Processes for Improved L-Valine Production","authors":"Caice Liang,&nbsp;Yong Wang,&nbsp;Shuqi Shi,&nbsp;Alan Yan,&nbsp;Qingguo Liu,&nbsp;Wei Liu,&nbsp;Hui Han,&nbsp;Wenlu Qi,&nbsp;Tianpeng Chen,&nbsp;Wenjun Sun,&nbsp;Yong Chen","doi":"10.1002/biot.70027","DOIUrl":"https://doi.org/10.1002/biot.70027","url":null,"abstract":"<div>\u0000 \u0000 <p>L-valine, an essential branched-chain amino acid, is widely used across various industrial sectors. Despite its significance, there is a scarcity of continuous fermentation methodologies, specifically for L-valine production. Biofilm-based Technologies face challenges in sustaining continuous L-valine production due to cell wall damage caused by L-valine sterilization. Here, we combined continuous fermentation principles with Biofilm-based technologies to develop a biofilm-based continuous fermentation (BCF) system, enabling sustained L-valine production. By analyzing the fermentation kinetics of free-cell fermentation (FCF), we optimized the L-valine titer in BCF to 60–70 g/L, achieving a yield of 0.44 g/g glucose—a 10% increase over FCF. Notably, reducing the controlled L-valine titer in BCF extended the fermentation time but simultaneously decreased both the yield and production rate. Conversely, shortening the fermentation time was associated with an increase in both yield and production rate. In summary, BCF significantly improved L-valine purity, yield from glucose, and production rate while reducing by-product formation.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Mechanism Underlying Enhanced Coal-to-Methane Conversion in Anaerobic Digestion With Betaine Supplementation","authors":"Zhiting Di,&nbsp;Yaya Wang,&nbsp;Enxi Zhang,&nbsp;Ye Zhang,&nbsp;Yuan Bao,&nbsp;Kaile Zhao,&nbsp;Zhigang Wang,&nbsp;Shihua He,&nbsp;Yanxin Xiang","doi":"10.1002/biot.70028","DOIUrl":"https://doi.org/10.1002/biot.70028","url":null,"abstract":"<div>\u0000 \u0000 <p>Anaerobic digestion (AD) of coal-to-methane technology represents a promising energy conversion method that not only reduces environmental pollution but also contributes to sustainable development. However, low methane conversion efficiency remains a major challenge. This study investigates the use of betaine, a green and environmentally friendly alkaloid, to enhance the AD process of coal. Our results demonstrate that betaine supplementation increases the relative abundance of key microbial populations, including <i>Petrimonas</i>, <i>Desulfitibacter</i>, <i>Methanoculleus</i>, and <i>Methanosarcina</i>, while also improving cell membrane permeability. Three-dimensional fluorescence characterization reveals elevated secretion of bacterial metabolites, leading to increased protein concentrations and enhanced enzymatic activity in the liquid phase. Moreover, the content of alkane compounds in the liquid phase increases, further confirming the enhanced conversion of coal to biological methane. In conclusion, betaine supplementation significantly improves coal AD efficiency, providing a novel approach to optimize coal fermentation and biological energy conversion.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bacillus Subtilis as an Excellent Microbial Treatment Agent for Environmental Pollution: A Review","authors":"Zhuoman Wang,&nbsp;Zhongyuan Li,&nbsp;Cuimei Gao,&nbsp;Zijian Jiang,&nbsp;Siqi Huang,&nbsp;Xin Li,&nbsp;Huilin Yang","doi":"10.1002/biot.70026","DOIUrl":"https://doi.org/10.1002/biot.70026","url":null,"abstract":"<div>\u0000 \u0000 <p>The use of microorganisms in environmental biotreatment is gaining attention, particularly <i>Bacillus subtilis (B. subtilis)</i>, recognized for its effectiveness in wastewater treatment and soil remediation. Its success stems from its diverse biological activities and adaptability, which improve environmental quality and ecological balance. This paper reviews the remediation capabilities and mechanisms of <i>B. subtilis</i>, focusing on its applications in water purification and soil pollution management. <i>B. subtilis</i> facilitates pollutant degradation and adsorption through enzyme production, organic acids, unique cell wall properties, and interactions with other microorganisms. The article addresses current challenges and future directions, emphasizing the need for enhanced cultivation, screening, and genetic engineering of functional strains. Understanding the interactions of these strains with other microorganisms and studying their ecological and toxicological impacts are essential for optimizing microbial remediation, providing both theoretical and practical foundations for bioremediation efforts.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875594","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Two-Step Adaptive Laboratory Evolution Enhances Osmotolerance in Engineered Escherichia coli for Improved Succinate Biosynthesis","authors":"Yanzhe Shang,&nbsp;Zhengtong Zhu,&nbsp;Junru Sun,&nbsp;Peng Fei,&nbsp;Yuanchan Luo,&nbsp;Hui Wu","doi":"10.1002/biot.70021","DOIUrl":"https://doi.org/10.1002/biot.70021","url":null,"abstract":"<div>\u0000 \u0000 <p>Succinic acid (SA) is a promising platform chemical with broad applications in agricultural, food, and pharmaceutical industries. Microbial production of SA using <i>Escherichia coli</i> typically requires alkaline neutralizers to maintain pH during fermentation, leading to elevated osmotic pressure that severely inhibits SA production. The strain ZZT215, evolved from AFP111 using two-step adaptive laboratory evolution (ALE) strategy, exhibited the improved Na⁺ tolerance and SA productivity without further genetic modification. In 5 L bioreactor fermentation, ZZT215 accumulated 87.02 g/L of SA with a productivity at 1.01 g/(L·h), representing 24.9% and 21.7% increases compared to the parent strain AFP111, respectively. Transcriptomic analysis revealed downregulated TCA cycle genes and upregulated ABC transporters, indicating metabolic adaptation to osmotic stress. These findings highlight the potential of multiple-step ALE for engineering robust microbial cell factories for SA and other high-value chemicals.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143831256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Generation of MCF-7 Spheroids in Polyethylene Glycol-Dextran Droplets for Cancer Niche Studies Using Aqueous Two-Phase System-3D Platforms","authors":"Karolina Chairez-Cantu,&nbsp;Mirna González-González,&nbsp;Marco Rito-Palomares","doi":"10.1002/biot.70014","DOIUrl":"https://doi.org/10.1002/biot.70014","url":null,"abstract":"<div>\u0000 \u0000 <p>Cell-based aqueous two-phase system (ATPS) applications involve the construction of 3D cultures encapsulating cells in polyethylene glycol (PEG)-dextran (DEX) droplets due to their low interfacial tension. This technology has enabled cell patterning in a controlled-defined shape giving rise to specific cell microenvironments. The present work aims to encapsulate MCF-7 cancer cells in ATPS droplets evaluating four different compositions of PEG-DEX, construction Strategies A (droplet added), B (immersed), or C (covered), and two cell densities (5000 and 10,000 cells/µL) during a 24 h incubation time using 96-well plates. The smallest cell-containing DEX droplets showed a mid-range value of 1.05 ± 0.33 mm using Strategy C whereas the largest of 1.46 ± 0.49 mm in Strategy B. However, Strategy A was associated with higher rates of circularity. After 24 h, MCF-7 spheroids were formed in all PEG-DEX systems showing a diameter size ranging from 0.10 to 0.65 mm. Their circularity was higher in low cell densities. Higher rates of cell viability were obtained in PEG 35,000–DEX 500,000 systems for a period of 4 days showing tumor-specific traits compared to 2D control. This work demonstrates practical strategies for 3D model construction for cancer niche for future drug screening studies using basic laboratory equipment.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143827083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of High-Speed Shaking on Oxygen Transfer in Shake Flasks","authors":"Andreas Schulte,&nbsp;Andreas Jordan,&nbsp;Wolf Klöckner,&nbsp;Mathias Schumacher,&nbsp;Burkhard Corves,&nbsp;Jochen Büchs","doi":"10.1002/biot.70013","DOIUrl":"https://doi.org/10.1002/biot.70013","url":null,"abstract":"<p>Shake flasks are predominantly used in screening and the early stages of biotechnological process development. However, oxygen-demanding processes cannot easily be performed in shake flasks, since the maximum oxygen transfer capacity is usually smaller than in stirred reactors. Studies during the last decades suggest that the shaking frequency is one of the most crucial cultivation parameters to sustainably increase oxygen supply in orbitally shaken bioreactors. In this study — for the first time — a prototype of a self-balancing orbital shaker was used, which is capable to be operated at up to 750 rpm shaking frequency at 25 mm shaking diameter and 600 rpm at 50 mm. <i>Kluyveromyces lactis</i> cultivations were monitored with a modified TOM system to measure the maximum oxygen transfer capacities (OTR_max) and corresponding <i>k_La</i> values. A maximum <i>k_La</i> value of 650 h⁻¹ (OTR_max = 135 mmol/L/h) was reached at 10 mL filling volume in a 250 mL shake flask made of glass with a hydrophilic surface property. This is an increase of about 50%, compared to current commercial orbital shakers. The new high-speed orbital shaker provides new possibilities for screening applications and process development. High-speed shaking for enhanced oxygen supply is particularly beneficial at 25 mm shaking diameter, rather than at 50 mm, minimizing the impact of the elevated centrifugal force on the shaking system.</p>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70013","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143827082","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":"Novel Approach Using Real-Time Dynamic Imaging Analysis to Monitor Cellular Apoptosis, Viability, and Cell Density in CHO Cell-Based Bioprocesses for Monoclonal Antibody Production","authors":"Claudia Corredor,&nbsp;Yuxiang Zhao,&nbsp;Suyang Wu,&nbsp;James K. Drennen,&nbsp;Carl Anderson","doi":"10.1002/biot.70018","DOIUrl":"https://doi.org/10.1002/biot.70018","url":null,"abstract":"<div>\u0000 \u0000 <p>We investigated an analytical method based on dynamic imaging analysis (DIA) for monitoring cellular apoptosis, viability, and cell density. Trypan blue and flow cytometry were used as reference methods. The DIA method showed results comparable to the traditional methods, with clear advantages (label-free and in-line real-time monitoring capability). The DIA method successfully traced the trajectory of cell death progress, detecting the onset of apoptosis earlier than the other methods.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143827084","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ta-Ag Coatings on TC4: A Strategy to Leverage Bioelectric Microenvironments for Enhanced Antibacterial Activity","authors":"Yuxin Gong,&nbsp;Xiang Liang,&nbsp;Le Bai,&nbsp;Ming Yu,&nbsp;Xin Yang,&nbsp;Chonghao Yao,&nbsp;Hao Cui,&nbsp;Linyang Xie,&nbsp;Bingheng Lu,&nbsp;Sijia Na,&nbsp;Guangbin Zhao,&nbsp;Junbo Tu,&nbsp;Fangfang Xu","doi":"10.1002/biot.70000","DOIUrl":"https://doi.org/10.1002/biot.70000","url":null,"abstract":"<p>Dental implant-related infections are serious complications after surgery that can results in loosening or even complete loss of the implant. Although endogenous electric fields (EEF) play an integral role in the human body, current methods involving external electrical stimulation are invasive and not suitable for clinical application. In this study, we using DC magnetron sputtering, investigates the effects of tantalum-silver (Ta-Ag) coatings on titanium alloy (TC4) surfaces, focusing on their potential to influence EEF that enhances antibacterial activity In this design, Ta-Ag configuration effectively increased the surface potential difference of TC4, and furthermore, promoting Ta/Ag ions release and reducing bacterial adhesion. The study concludes that the Ta-Ag coating, particularly the TT/A implant, promotes a stable EEF, enhancing the long-term antibacterial and osteogenic properties of implants. This work provides a promising strategy for developing advanced implant materials with improved clinical efficacy.</p>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"20 4","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/biot.70000","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143827085","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}