Biotechnology and Bioengineering最新文献

{"title":"Physiological Response of Penicillium chrysogenum to Mimicked Local and Global Perturbations of Substrate and Dissolved Oxygen Gradients at Industrial-Scale","authors":"Yining Chen, Cees Haringa, Zejian Wang, Yingping Zhuang, Guan Wang","doi":"10.1002/bit.28968","DOIUrl":"https://doi.org/10.1002/bit.28968","url":null,"abstract":"Industrial-scale microbial fermentation processes often face limitations in mixing and mass transfer, leading to the formation of environmental gradients within the bioreactor. These gradients expose microbes to heterogeneous conditions over time and space. In this study, we evaluated the effects of combined substrate and dissolved oxygen (DO) gradients on the metabolic response of <i>Penicillium chrysogenum</i> at an industrial scale. Three representative heterogeneous environments were simulated in scale-down systems: (1) feed inlet (high glucose, low oxygen (HGLO): C_S &gt; 20 mM, DO &lt; 0.012 mM), (2) aeration inlet (high oxygen, low glucose (HOLG): C_S &lt; 0.8 mM, DO &gt; 0.2 mM), and (3) global environment (periodic 360 s fluctuation cycle with 45 s of HGLO and 75 s of HOLG conditions). Results showed that prolonged exposure to feed inlet conditions led to a complete loss of penicillin production capacity, accompanied by significant excretion of intracellular metabolites, and this effect was largely irreversible. While, cells randomly walking under the top impeller zone did not lose production capacity but showed signs of premature degeneration due to increased energy demand. When exposed to the global environment, cells finely tuned their metabolism in a periodical manner, with nearly a 50% loss of penicillin productivity. In summary, substrate gradients alone did not cause irreversible effects, but large substrate gradients contributed to reduced productivity. Oxygen gradients, however, not only reduced production but also caused irreversible cellular damage. These findings provide valuable insights for developing scale-up criteria and strain engineering strategies aimed at improving large-scale culture performance.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"26 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143608706","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 Printable Self-Healing Mineralized Hydrogels Loaded With Diclofenac Sodium: In Vitro and In Vivo Assessment","authors":"Nachiketa Palit, Purushottam Suryavanshi, Subham Banerjee","doi":"10.1002/bit.28973","DOIUrl":"https://doi.org/10.1002/bit.28973","url":null,"abstract":"The use of self-healing mineralized hydrogels in 3D printing has demonstrated significant advantages, including enhanced printing accuracy and the ability to maintain high shape fidelity throughout the printing process. After conducting an initial optimization study, we incorporated our self-healing mineralized hydrogel into semi-solid extrusion-based 3D printing to print diclofenac-loaded oral films. The dependence of the print speed on the nature of the material was established by varying the print speed. The process of optimizing the print speed was conducted using a blank hydrogel, which involved analyzing specific parameters, such as printing accuracy and the percentage of pore area under sizing. The results demonstrated that 2 mm/sec print speed showed a higher printing accuracy of 98.13% and pore area under-sizing value of 41.31%. Interestingly, the viscosity of the hydrogel increased from 5.30 to 133 PaS upon addition of the drug. The percentage pore area under sizing also decreased from 41.31% to 11.48% as the drug loading was increased from 0% to 3% <i>w</i>/<i>w</i>. The in vitro drug release study demonstrated that the 3% <i>w</i>/<i>w</i> diclofenac sodium-loaded oral films printed at 2 mm/sec exhibited a faster release profile. Furthermore, considerable bioavailability of diclofenac sodium (DS) was achieved from the 3D-printed oral films during the in vivo study. These results can be effectively used to develop a drug delivery system that can release medications accurately and consistently, either in a targeted area or systemically.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"68 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143598861","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 122, Number 4, April 2025","authors":"","doi":"10.1002/bit.28744","DOIUrl":"https://doi.org/10.1002/bit.28744","url":null,"abstract":"","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 4","pages":"745-748"},"PeriodicalIF":3.5,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.28744","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143594962","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":"Corrigendum to “High-Throughput Investigation of Endothelial-to-Mesenchymal Transformation (EndMT) With Combinatorial Cellular Microarrays”","authors":"","doi":"10.1002/bit.28970","DOIUrl":"https://doi.org/10.1002/bit.28970","url":null,"abstract":"<p>Zongjie W., B. Calpe, J. Zerdani, Y. Lee, J. Oh, H. Bae, A. Khademhosseini, and K. Kim. 2016. “High-Throughput Investigation of Endothelial-to-Mesenchymal Transformation (EndMT) With Combinatorial Cellular Microarrays.” <i>Biotechnology and Bioengineering</i>. 113, no. 7: 1403–1412.</p>\u0000<p>In Figure 3A, LM+C4 and C4 were mistakenly shown as identical due to an error in which the C4 array image was montaged from LM+C4 data. To correct this, the authors generated a new C4 array image using the correct C4 data. The corrected figure is shown below. This update does not affect the interpretation of the data or the study's conclusions.</p>\u0000<figure><picture>\u0000<source media=\"(min-width: 1650px)\" srcset=\"/cms/asset/9c17b85b-a4d5-4c71-add6-5d21e00698a3/bit28970-fig-0001-m.jpg\"/><img alt=\"Details are in the caption following the image\" data-lg-src=\"/cms/asset/9c17b85b-a4d5-4c71-add6-5d21e00698a3/bit28970-fig-0001-m.jpg\" loading=\"lazy\" src=\"/cms/asset/38bef1dd-9333-49ea-8b34-1617104b7c63/bit28970-fig-0001-m.png\" title=\"Details are in the caption following the image\"/></picture><figcaption>\u0000<div><strong>Figure 3<span style=\"font-weight:normal\"></span></strong><div>Open in figure viewer<i aria-hidden=\"true\"></i><span>PowerPoint</span></div>\u0000</div>\u0000<div> </div>\u0000</figcaption>\u0000</figure>\u0000<p>We apologize for this error.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"29 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143598817","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":"Synergistic Effects of a Novel Multifunctional Bionic Scaffold and Electrical Stimulation Promote Bone Tissue Regeneration.","authors":"Yingxin Zhang, Huanyan Dai, Xi Li, Zhiyan Wu, Zhimin Xu, Peng Liu, Bing Han","doi":"10.1002/bit.28964","DOIUrl":"https://doi.org/10.1002/bit.28964","url":null,"abstract":"<p><p>Electrical stimulation (ES) can effectively regulate cell behavior and promote bone tissue regeneration, and conductive biomaterials can further enhance this effect by enhancing the conduction of electrical signals between cells. In this study, poly(lactic-co-glycolic acid) (PLGA) and poly(l-lactide)-aniline pentamer triblock copolymer (PAP) were used as raw materials to prepare a conductive bionic scaffold (PLGA/PAP). Subsequently, bone morphogenetic protein 2 mimetic peptide containing a DOPA tag (DBMP2MP) was loaded on the scaffold surface. The prepared scaffold (DBMP2MP@PLGA/PAP) had a porosity of 79.17% and a porous structure similar to that of natural cancellous bone. After PAP was added, the mechanical strength and electrical conductivity of the scaffold were increased to 2.79 ± 0.1 kPa and 1.29 ± 0.023 × 10^-6 s/cm. The addition of DBMP2MP significantly improved the hydrophilicity of the scaffold material, and the contact Angle of the scaffold material decreased from 102.45 ± 7.67° to 30.36 ± 5.25°. At the same time, DBMP2MP and scaffold surface bonding ability increased by two times compared with commercial BMP2. The polypeptide DBMP2MP can bind to the surface of scaffolds and exhibit long-lasting biological effects. In vitro cell experiments revealed that the DBMP2MP@PLGA/PAP scaffold could significantly promote the proliferation and adhesion of MC3T3-E1 cells and that the combination of DBMP2MP@PLGA/PAP with pulsed ES could further synergistically induce cell mineralization and osteogenic differentiation. The results of the rabbit radius defect experiments revealed that grafting the DBMP2MP@PLGA/PAP scaffold at the defect site significantly promoted the formation of new bone and collagen fibers. When the DBMP2MP@PLGA/PAP scaffold was combined with ES, the regeneration rate of bone tissue further improved, and the newborn collagen tissue is close to normal bone collagen. Therefore, this bionic scaffold with excellent electrical and biological activity shows considerable potential in the field of bone defect repair.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143596152","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":"An Overview of Virus-Free Protein Expression in Insect Cells: A Mode of Rapid Manufacturing Platform for Therapeutic Protein and Virus-Like-Particles.","authors":"Jagadeesh Mahadevan, Kishalay Mitra, Lopamudra Giri","doi":"10.1002/bit.28961","DOIUrl":"https://doi.org/10.1002/bit.28961","url":null,"abstract":"<p><p>Production of therapeutic proteins, antibodies, and virus-like particles (VLP) using baculovirus expression systems (BEVS) has been explored for decades. However, we have realized an urgent need for accelerated production of recombinant proteins and VLPs to address critical situations in recent scenarios. In contrast to BEVSs, the virus-free method is significantly shorter as it bypasses the time-consuming process of infectivity monitoring and virus amplification. Moreover, in the virus-free method, complex steps of protein separation can be eliminated to ease downstream processing. Hence, we present a detailed review of the recent techniques for expressing recombinant proteins, therapeutics, and VLP in insect cells using virus-free methods. First, we focus on the specific methodologies used to optimize virus-free transfection. Here, we provide insight into the interplay between crucial factors, including concentration of transfection reagent, seeding density, and medium temperature. Secondly, we provide a structured review of the novel transfection reagents used for transient and stable transfection. Thirdly, we performed an assessment of the cell lines and plasmids used for virus-free expression and their evaluation based on corresponding protein yield. Finally, we provide the recent advancement in scaling up the transfection process from the shaker flask to the bioreactor level to achieve better yield. Various virus-free expression methodologies presented in this article are essential for evaluating the transfection processes toward improving protein yield. The readers can also use the information to design experiments and optimize process parameters for bioreactor operation.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143596134","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":"High-Efficiency PET Degradation With a Duo-Enzyme System Immobilized on Magnetic Nanoparticles","authors":"Siddhi Kotnis, Siddhant Gulati, Qing Sun","doi":"10.1002/bit.28963","DOIUrl":"https://doi.org/10.1002/bit.28963","url":null,"abstract":"The widespread consumption of PET worldwide has necessitated the search for environment-friendly methods for PET degradation and recycling. Among these methods, biodegradation stands out as a promising approach for recycling PET. The discovery of duo enzyme system PETase and MHETase in 2016, along with their engineered variants, has demonstrated significant potential in breaking down PET. Previous studies have also demonstrated that the activity of the enzyme PETase increases when it is immobilized on nanoparticles. To achieve highly efficient and complete PET depolymerization, we immobilized both FAST-PETase and MHETase at a specific ratio on magnetic nanoparticles. This immobilization resulted in a 2.5-fold increase in product release compared with free enzymes. Additionally, we achieved reusability and enhanced stability of the enzyme bioconjugates.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"49 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143582968","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":"Engineered Lactobacillus rhamnosus Producing 3-Hydroxybutyrate: A Dual-Action Therapeutic Strategy for Colon Cancer Cachexia","authors":"Xu Qiu, Zhiyun Zou, Taijie Lin, Chenyun Guo, Donghai Lin","doi":"10.1002/bit.28972","DOIUrl":"https://doi.org/10.1002/bit.28972","url":null,"abstract":"3-hydroxybutyrate (3-HB), an essential endogenous metabolite, shows significant therapeutic potential in several disease contexts. However, its clinical application has been hampered by limitations, such as adverse effects on the gut microbiota. This study introduces a genetically engineered strain of <i>Lactobacillus rhamnosus</i> GG (LGGK) that integrates the benefits of 3-HB production with the probiotic properties of LGG. Using a murine colon cancer cachexia (CAC) model, LGGK supplementation significantly improved survival, reduced tumor progression, and alleviated muscle wasting. LGGK restored gut microbial diversity, increased the abundance of beneficial bacteria, and increased the production of short-chain fatty acids while reducing harmful microbial populations. In addition, LGGK supplementation demonstrated anti-inflammatory effects, effectively reducing elevated pro-inflammatory cytokines in serum and skeletal muscle. These findings highlight LGGK as a dual-action therapeutic approach that utilizes the metabolic benefits of 3-HB and the gut-modulating properties of LGG. This innovation offers a promising strategy for the treatment of CAC and potentially other metabolic and inflammatory disorders, and highlights the potential of engineered probiotics in advanced therapeutic applications.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"53 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143575227","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":"Self-Healable Hydrogel for Regression of Liver Fibrosis","authors":"Yu-Chai Tai, Wei-Rong Yin, Kai-Yi Cheng, Xin-Yu Chou, Yong-Heng Lin, Shan-hui Hsu, Yung-Te Hou","doi":"10.1002/bit.28966","DOIUrl":"https://doi.org/10.1002/bit.28966","url":null,"abstract":"Liver fibrosis is considered as a wound healing process in the presence of chronic hepatic injury. A hydrogel (CPDP) based on chitosan–phenol that undergoes fast gelling and owns self-healing and injectable properties was investigated for the effect on regression of liver fibrosis. For the purpose, we established both in vitro and in vivo liver fibrosis models and implanted CPDP hydrogel into the injured liver. The CPDP hydrogel not only provided a suitable microenvironment for hepatocyte spheroids, but also demonstrated a potential for the hepatocyte spheroid-embedded system to mimic the liver tissue in vitro. Furthermore, the urea synthesis of injured hepatocytes cultured on hepatocyte spheroid-embedded CPDP hydrogel was 1.12 times higher than that on hepatocyte spheroid-embedded collagen hydrogel after 7 days of culture, indicating that CPDP hydrogel effectively rescued hepatic function in the injured hepatocytes. Moreover, the hepatic injury was alleviated with improved hepatic function in the liver fibrosis model in vivo. A reduction of approximately 28% in serum AST/ALT ratios and a 70% decrease in the fibrotic area suggested the regression of liver fibrosis after 2 weeks of CPDP hydrogel administration. These findings suggest that CPDP hydrogel holds promise for applications in liver tissue engineering.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"12 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143569574","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":"Engineering Affibody Binders to Death Receptor 5 and Tumor Necrosis Factor Receptor 1 With Improved Stability","authors":"Gregory H. Nielsen, Jonathan N. Sachs, Benjamin J. Hackel","doi":"10.1002/bit.28954","DOIUrl":"https://doi.org/10.1002/bit.28954","url":null,"abstract":"Protein developability is an important, yet often overlooked, aspect of protein discovery campaigns that is a key driver of utility. Recent advances have improved developability screening capacity, making it an increasingly viable option in early-stage discovery. Here, we engineered one component of developability, stability, of two affibody proteins—one that targets death receptor 5 and another that targets tumor necrosis factor receptor 1—previously evolved to bind receptor and non-competitively inhibit signaling via conformational modulation. Starting from an error-prone PCR library of each affibody, variants were screened via yeast surface display binder selections, including depletion of non-specific binders, followed by developability assessment using the on-yeast protease and yeast display level assays. Multiplex deep sequencing identified variants for further evaluation. Purified variants exhibited elevated stability—8°C to 14°C increase in T_m,app—with maintained 1–2 nM affinity for the TNFR1 affibody and 30-fold improvement in the DR5 affibody affinity to 0.8 nM.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"53 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143560759","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}