Biotechnology and Bioengineering最新文献

{"title":"Derivation of an Upscaled Model for Xylitol Production With Immobilized Microorganisms","authors":"Rolando Zenteno‐Catemaxca, Roel Hernandez‐Rodriguez, Epifanio Morales‐Zárate, Eliseo Hernandez‐Martinez","doi":"10.1002/bit.28946","DOIUrl":"https://doi.org/10.1002/bit.28946","url":null,"abstract":"Xylitol, a polyalcohol with anticariogenic properties, finds broad applications in different sectors. Research into new production methods has highlighted the biochemical route using immobilized microorganisms. However, challenges remain in optimizing operating conditions and understanding mass transport and biochemical reactions. Different macroscopic models have been proposed to address these challenges. Nevertheless, those models do not consider porous particles' microstructure and biofilms' formation within them, which can determine the macroscopic performance of the process due to its hierarchical nature. In this work, we derive two macroscopic models for the mass transport and reaction of the xylitol production process with immobilized microorganisms in porous particles. Such models are derived from microscopic ones using the volume averaging method, resulting in both two‐equation and one‐equation models, written in terms of effective medium coefficients. These latter are predicted by solving ancillary problems in representative 2D unit cells of the immobilization particles, incorporating their microstructural information. Besides, kinetic parameters are estimated through kinetic fitting using experimental data from the literature. Models' accuracy is assessed by comparing them with pore‐scale simulations and experimental observations of xylitol production from sugarcane bagasse at the laboratory scale, finding good agreement. Finally, our results are compared with a macroscopic model reported in the literature, and similar predictions are found. However, unlike the reported model, the one derived here improves the modeling of the process since the effective coefficients do not need to be calculated using empirical correlations or estimators.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"1 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143426990","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":"Zeolitic Imidazole Framework-8 Nanoparticles as an Alternative to Freund's Adjuvant for Klebsiella pneumoniae Recombinant Protein Vaccine","authors":"Gaowei Hu, Chunli Hong, Yingjie Miao, Wenji Wang, Longfei Yin, Xi Luo, Yongqian Fu","doi":"10.1002/bit.28944","DOIUrl":"https://doi.org/10.1002/bit.28944","url":null,"abstract":"Vaccination represents a promising approach to combat resistant <i>Klebsiella pneumoniae</i> (KP). However, there is currently no licensed vaccine in the veterinary field. Outer membrane proteins have been proven to possess good immunogenicity, but Freund's adjuvant, which is commonly used to administer protein vaccines, has limitations such as a complicated formulation process as well as a tendency to cause pain and inflammation in animals. Here, we prepared a nano-vaccine based on zeolitic imidazolate framework-8 (ZIF-8)-encapsulated outer membrane protein PhoE and evaluated its efficiency in enhancing humoral and cellular immune responses in BALB/c mice. ZIF-8 nanoparticles rapidly delivered the protein antigen into dendritic cells and successfully activated them. In addition, significantly higher IgG antibody titers, cytokine levels, and splenocyte proliferation indices were founded in mice subcutaneously immunized with PhoE@ZIF-8 than in those receiving free PhoE alone. In a BALB/c mouse model, PhoE@ZIF-8 elicited a strong immune response with improved prophylactic efficacy against KP that was similar to the Freund's adjuvant-formulated vaccine. Based on the superiority of this nano-vaccine with good biocompatibility, inexpensive preparation and higher efficiency of delivering antigen into cells, ZIF-8 can serve as a promising replacement for Freund's adjuvant in research, with a prospective usage for vaccines against bacterial pathogens in the veterinary field.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"9 30 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143393364","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":"Development of Methods to Produce SARS CoV-2 Virus-Like Particles at Scale","authors":"Melissa A. Edeling, Linda Earnest, Julio Carrera Montoya, Ashley Huey Yiing Yap, Jamie Mumford, Jason Roberts, Chinn Yi Wong, Dhiraj Hans, Joseph Grima, Nicole Bisset, Jesse Bodle, Steven Rockman, Joseph Torresi","doi":"10.1002/bit.28937","DOIUrl":"https://doi.org/10.1002/bit.28937","url":null,"abstract":"The devastating global toll precipitated by the SARS CoV-2 outbreak and the profound impact of vaccines in stemming that outbreak has established the need for molecular platforms capable of rapidly delivering effective, safe and accessible medical interventions in pandemic preparedness. We describe a simple, efficient and adaptable process to produce SARS CoV-2 virus-like particles (VLPs) that can be readily scaled for manufacturing. A rapid but gentle method of tangential flow filtration using a 100 kDa semi-permeable membrane concentrates and buffer exchanges 0.5 L of SARS CoV-2 VLP containing supernatant into low salt and optimal pH for anion exchange chromatography. VLPs are washed, eluted under high salt, dialyzed into physiological buffer, sterile filtered and aliquoted for storage at –80°C. Purification is completed in less than 2 days. A simple quality control process includes Western blot for coincident detection of Spike, Membrane and Envelope protein as a proxy for intact VLP, ELISA to detect conformationally sensitive Spike using readily available anti-Spike and/or anti-RBD antibodies, and negative stain and immunogold electron microscopy to validate particulate, Spike crowned VLPs. This process to produce SARS CoV-2 VLPs for preclinical studies serves as a roadmap for preparation of more distantly related VLPs for pandemic preparedness.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"18 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143393362","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":"Assessing the Adjuvant Potential of Chinese Hamster Ovary Host Cell Proteins Using an In Vitro Dendritic Cell Assay","authors":"Sherin Panikulam, Hannah Morgan, Michael Gutknecht, Anette Karle, Atchaya Rajaratnam, Jennifer Muntwyler, Oliver Anderka, Nicolas Lebesgue, Thomas K. Villiger","doi":"10.1002/bit.28950","DOIUrl":"https://doi.org/10.1002/bit.28950","url":null,"abstract":"Host cell proteins (HCPs) are process-related impurities of therapeutic protein production and may affect product quality or patient safety. In clinical trials, certain HCPs (e.g., PLBL2 or CCL2) that co-purify with the therapeutic protein have been associated with immune reactions in patients. In this study, we examined the adjuvant potential of six commonly detected HCPs from CHO cells (PRDX1, S100A4, PLBL2, CCL2, CLU, and YWHAE) using an in vitro dendritic cell (DC) maturation assay. Recombinant HCPs were expressed in CHO cells to mimic manufacturing conditions. PRDX1, S100A4, and PLBL2 caused a slight increase in the expression of maturation markers on DCs, while YWHAE, CLU, and CCL2 did not. Interestingly, CLU and CCL2 reduced the DC maturation induced by rituximab. In addition, we observed that process parameters such as elution conditions during chromatographic purification can influence HCP aggregation, which in turn can mask or enhance the intrinsic adjuvant potential of an HCP. These findings not only provide initial insights into the adjuvant potential of individual HCPs but also indicate that the quantity as well as the degree of aggregation of HCPs might influence adjuvanticity.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"4 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143385828","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 3, March 2025","authors":"","doi":"10.1002/bit.28743","DOIUrl":"10.1002/bit.28743","url":null,"abstract":"","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 3","pages":"453-456"},"PeriodicalIF":3.5,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.28743","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143375807","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":"Enhanced Natural Killer Cell Proliferation by Stress-Induced Feeder Cells","authors":"Donghyun Lee, Myeongkwan Song, Soonjo Kwon","doi":"10.1002/bit.28951","DOIUrl":"https://doi.org/10.1002/bit.28951","url":null,"abstract":"Natural killer (NK) cells, integral to the innate immune system, are notable in cell therapies because of their applicability in allogeneic treatments, distinguishing them from T cells typically employed in conventional cell therapies. However, their limited half-life (proliferative capability) poses a challenge for therapy. The limited half-life creates difficulties in obtaining a sufficient number of cells for in vitro adoptive therapy. Gene modification is commonly employed to address this limitation. However, due to concerns such as genetic instability and unintended gene expression, its suitability for long-term cultivation is uncertain. Consequently, safer alternatives are needed. We aimed to promote NK cell proliferation through feeder cells rather than genetic modification. These cells are designed to interact with NK cells without adverse effects, aiming to promote NK cell proliferation more safely. In our study, during the tailoring of feeder cells, we excluded genetic modification and instead applied chemical-based extracellular stress. The extracellular stress applied consisted of hypoxia and cytochalasin D. By treating the feeder cells with these stressors, we were able to inhibit feeder cell proliferation, enabling them to function more efficiently as feeder cells. Furthermore, we observed that the feeder cells subjected to extracellular stress exhibited upregulated expression of 4-1BBL, which enhances the 4-1BB/4-1BBL interaction with NK cells. The upregulated 4-1BBL binds to 4-1BB on the surface of NK cells, promoting their proliferation. Additionally, following coculture with feeder cells exposed to extracellular stress, we observed an upregulation of CD56 expression on the surface of NK cells. These CD56^bright NK cells influence NK cell proliferation through enhanced cytokine release. We further validated this process under dynamic conditions where shear stress is applied, demonstrating that the feeder cell-mediated enhancement of NK cell proliferation is applicable under dynamic conditions such as those found in bioreactors.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"78 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143385827","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":"One-Pot Hetero-Di-C-Glycosylation of the Natural Polyphenol Phloretin by a Single C-Glycosyltransferase With Broad Sugar Substrate Specificity","authors":"Tuo Li, Annika J. E. Borg, Leo Krammer, Rolf Breinbauer, Bernd Nidetzky","doi":"10.1002/bit.28948","DOIUrl":"https://doi.org/10.1002/bit.28948","url":null,"abstract":"The structural motif of hetero-di-<i>C</i>-glycosyl compound is prominent in plant polyphenol natural products and involves two different glycosyl residues (e.g., β-<span>d</span>-glucosyl, β-<span>d</span>-xylosyl) attached to carbons of the same phenolic ring. Polyphenol hetero-di-<i>C</i>-glycosides attract attention as specialized ingredients of herbal medicines and their tailored synthesis by enzymatic <i>C</i>-glycosylation is promising to overcome limitations of low natural availability and to expand molecular diversity to new-to-nature glycoside structures. However, installing these di-<i>C</i>-glycoside structures with synthetic precision and efficiency is challenging. Here we have characterized the syntheses of <i>C</i>-β-galactosyl-<i>C</i>-β-glucosyl and <i>C</i>-β-glucosyl-<i>C</i>-β-xylosyl structures on the phloroglucinol ring of the natural polyphenol phloretin, using kumquat (<i>Fortunella crassifolia</i>) <i>C-</i>glycosyltransferase (<i>Fc</i>CGT). The <i>Fc</i>CGT uses uridine 5'-diphosphate (UDP)-galactose (5 mU/mg) and UDP-xylose (0.3 U/mg) at lower activity than UDP-glucose (3 U/mg). The 3'-<i>C</i>-β-glucoside (nothofagin) is ~10-fold less reactive than non-glycosylated phloretin with all UDP-sugars, suggesting the practical order of hetero-di-<i>C</i>-glycosylation as <i>C</i>-galactosylation or <i>C</i>-xylosylation of phloretin followed by <i>C</i>-glucosylation of the resulting mono-<i>C</i>-glycoside. Each <i>C</i>-glycosylation performed in the presence of twofold excess of UDP-sugar proceeds to completion and appears to be effectively irreversible, as evidenced by the absence of glycosyl residue exchange at extended reaction times. Synthesis of <i>C</i>-β-glucosyl-<i>C</i>-β-xylosyl phloretin is shown at 10 mM concentration in quantitative conversion using cascade reaction of <i>Fc</i>CGT and UDP-xylose synthase, allowing for in situ formation of UDP-xylose from the more expedient donor substrate UDP-glucuronic acid. The desired di-<i>C</i>-glycoside with Xyl or Gal was obtained as a single product of the synthesis and its structure was confirmed by NMR.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"9 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143257834","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":"Enhancing Biomass Productivity by Forecast-Informed Pond Operations","authors":"Hongxiang Yan, Mark S. Wigmosta, Ning Sun, Song Gao, Michael H. Huesemann","doi":"10.1002/bit.28952","DOIUrl":"https://doi.org/10.1002/bit.28952","url":null,"abstract":"Microalgal cultivation for biofuels and proteins holds significant promise but faces challenges in achieving economically viable biomass productivity under variable environmental conditions. This study introduces a forecast-informed pond operation (FIPO) system that uses numerical weather prediction (NWP) ensemble forecasts and the biomass assessment tool (BAT) to optimize daily dilution rates for enhanced biomass production. In contrast to the current practice, where fixed dilution rates are based on operator experience, the FIPO system determines the optimal dilution rate based on future weather forecasts and biomass growth conditions. Our experiments validate the effectiveness of FIPO in both short- and long-term growth scenarios. In short-term experiments, FIPO increased biomass production by 21.3% compared to batch growth and 7.4% over fixed dilution (60% every 3 days) operations. The NWP forecast-informed operations achieved biomass production nearly identical to that using perfect weather forecasts, highlighting the accuracy of current NWP forecasts for guiding pond operations. In long-term experiments, FIPO resulted in biomass production increases of 13.3% and 17.8% compared to two fixed dilution rates (60% every 3 days and 20% daily). These findings underscore the viability of using NWP forecasts to optimize microalgal cultivation systems. By adjusting daily dilution rates in response to forecasted weather, operators can achieve higher biomass yields and mitigate risks associated with environmental variability. This study provides a foundation for future research and practical applications in commercial-scale microalgal production.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"8 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143257833","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":"Systematic Development of a Detergent Toolbox as an Alternative to Triton X-100","authors":"Varsha Yadav, Mary Lunson, Hannah Shore, Jean Aucamp","doi":"10.1002/bit.28947","DOIUrl":"https://doi.org/10.1002/bit.28947","url":null,"abstract":"Detergents are routinely included in protein purification processes to inactivate enveloped viruses that may arise from adventitious or endogenous contamination. The detergent Triton X-100 (TX-100) has been widely used as part of the production process for therapeutic proteins. However, recent ecological studies indicate that TX-100 and its metabolites detrimentally impact aquatic organisms, thus alternative detergents for viral inactivation are required. The overall aim of this study was to identify one or more detergents that are a suitable replacement for TX-100 in the viral inactivation step. In stage one, 16 potential alternatives were identified and screened against TX-100 using multiple criteria such as solubility, feasibility of virus inactivation, critical micelle concentration, and storage conditions. The multi-criteria decision analysis (MCDA) methodology was used to identify four candidates for the second stage assessment. In stage two, a detailed evaluation was undertaken and two candidates C16-AO, and C11/15-sEO9, were found to be practical alternatives to TX-100 for use in protein therapeutic production processes for inactivating enveloped viruses. In addition, C13-EO8 demonstrated good viral inactivation capability and warrants further investigation in detergent clearance and impact on product quality.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"40 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143192638","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":"Construction of a CRISPR-Cas9-Based Genetic Editing Tool for Serratia marcescens Using a Stationary Phase Promoter and Its Application in Putrescine Production","authors":"Linbo Gou, Di Liu, Tai-Ping Fan, Huaxiang Deng, Yujie Cai","doi":"10.1002/bit.28949","DOIUrl":"https://doi.org/10.1002/bit.28949","url":null,"abstract":"Putrescine plays a significant role in green food production and agriculture by promoting plant growth and enhancing crop quality. Its application reduces the reliance on chemical fertilizers and pesticides, thereby supporting the advancement of sustainable agricultural practices. This study achieved efficient production of putrescine in <i>Serratia marcescens</i>. <i>S. marcescens</i> has been extensively used to synthesize antimicrobial substances and express proteins, but its application has been limited by the lack of efficient genome-editing tools. This study presents a CRISPR-Cas9-based tool for gene editing in <i>S. marcescens</i>. A dual-plasmid system was constructed, incorporating an editing template into the plasmid pEdit with target-specific sgRNA. A stationary phase promoter was used to express Cas9 from <i>Streptococcus pyogenes</i> protein, avoiding the need for additional inducers and ensuring efficient one-step gene knockout and integration. The tool demonstrated over 80% editing efficiency across various <i>S. marcescens</i> strains and enabled successful single-base mutations. Using this tool, we enhanced putrescine production in <i>S. marcescens</i> HBQA7, optimizing the expression of ornithine decarboxylase from <i>Clostridium aceticum</i> DSM1496 with the P2 promoter and identifying the optimal integration site. Putrescine production reached 8.46 g/L within 48 h. This study significantly advances <i>S. marcescens</i> gene editing and metabolic engineering.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"55 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143192639","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}