Engineering in Life Sciences最新文献

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Citrate Supplementation Modulates Medium Viscosity and Poly-γ-Glutamic Acid Synthesis by Engineered B. subtilis 168 柠檬酸盐补充对工程枯草芽孢杆菌培养基粘度和聚γ-谷氨酸合成的调节
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-03-04 DOI: 10.1002/elsc.70009
Frederik Völker, Kyra Hoffmann, Birthe Halmschlag, Sandra Maaß, Jochen Büchs, Lars M. Blank
{"title":"Citrate Supplementation Modulates Medium Viscosity and Poly-γ-Glutamic Acid Synthesis by Engineered B. subtilis 168","authors":"Frederik Völker,&nbsp;Kyra Hoffmann,&nbsp;Birthe Halmschlag,&nbsp;Sandra Maaß,&nbsp;Jochen Büchs,&nbsp;Lars M. Blank","doi":"10.1002/elsc.70009","DOIUrl":"https://doi.org/10.1002/elsc.70009","url":null,"abstract":"<p>The industrially attractive biopolymer poly-γ-glutamic acid (γ-PGA) is commonly produced by species of the genus <i>Bacillus</i> by co-feeding different carbon- and nitrogen-sources. Recent studies have highlighted the pivotal role of co-metabolization of a rapidly degradable carbon source such as glycerol together with citrate for γ-PGA production, independently fueling biomass generation as well as tricarboxylic acid (TCA) cycle precursor supply. With this study, we report that the sole presence of citrate in the production medium greatly influences growth behavior, γ-PGA production, and the viscosity of microbial cultures during biopolymer synthesis. Independent of the citrate concentration in the medium, only minor amounts of citrate were imported by <i>B. subtilis</i> 168 in the presence of glycerol due to carbon catabolite repression. However, a high citrate concentration resulted in a 6-fold increase in γ-PGA titer compared to low exogenous citrate levels. Data suggests that citrate was not used as a precursor in γ-PGA synthesis but rather influenced the fate of imported glutamate. The citrate concentration also affected medium viscosity as depletion resulted in a remarkable spike in culture broth viscosity. Additionally, cellular proteome analysis at different levels of citrate availability revealed significant changes in protein abundance involved in motility and fatty acid degradation.</p><p><i>Practical Application:</i> This research provides critical insights into optimizing γ-PGA production in <i>Bacillus subtilis</i>, particularly by using citrate supplementation to control medium viscosity and improve production yields. The study reveals that citrate not only plays a role in controlling viscosity but also influences intracellular glutamate metabolism, a key factor for γ-PGA synthesis. Citrate interacts with divalent cations such as Mg<sup>2+</sup> and Ca<sup>2+</sup>, reducing electrostatic interactions and thus decreasing viscosity in the medium. Additionally, while citrate uptake is limited due to carbon catabolite repression (CCR), even the minimal presence of citrate impacts growth and production. The findings suggest that citrate may trigger unexplored regulatory mechanisms affecting glutamate utilization. Their understanding opens new avenues for industrial optimization, which focus on enhancing glutamate synthesis pathways and exploring novel citrate-sensing mechanisms. Overall, this research lays the groundwork for improving the efficiency and consistency of γ-PGA production by fine-tuning media components and understanding their metabolic effects.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 3","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.70009","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143554595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Cracks Repairing and Resistance to Water Penetration Properties of Microbial Self-Healing Cement 微生物自愈水泥的裂缝修复及抗水渗透性能
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-02-28 DOI: 10.1002/elsc.70010
Luo Liu, Youxi Li, Jianrong Song, Junlai Zhou, Weijian Yi, Yangyang Ge, Kewei Gao
{"title":"Cracks Repairing and Resistance to Water Penetration Properties of Microbial Self-Healing Cement","authors":"Luo Liu,&nbsp;Youxi Li,&nbsp;Jianrong Song,&nbsp;Junlai Zhou,&nbsp;Weijian Yi,&nbsp;Yangyang Ge,&nbsp;Kewei Gao","doi":"10.1002/elsc.70010","DOIUrl":"https://doi.org/10.1002/elsc.70010","url":null,"abstract":"<p>This study focuses on applying microbial self-healing cement in repairing cracks in cement-based materials and enhancing its resistance to water penetration performance. Traditional cement is susceptible to environmental influences, leading to the formation of microcracks and a reduction in durability. This research used <i>Bacillus pseudofirmus</i> to prepare microcapsules through sodium alginate gelation technology. We mixed microcapsules into the cement. The results indicate that the microbial self-healing cement, with a 1% self-healing agent added, increased its resistance to water penetration ability by 29.2% after 28 days. This improvement rose to 39.3% after 84 days. Additionally, we used the embedded needle method to make mortar blocks through microcracks, mimicking the cracks found in real cement. The self-healing effect of the microcapsules was especially noticeable for cracks under 0.3 mm in diameter, compared to the commonly used commercial crystallization penetration technology. This is attributed to the crystalline bodies formed by the self-healing agent in the microcapsules blocking the cracks and preventing water penetration. This study provides an environmentally friendly solution for the repair of cracks in cement-based materials using microbial self-healing technology and lays the foundation for improving the repair efficiency and durability and exploring stability and reliability in the future.</p><p><i>Practical Application:</i> This study investigated the application of microbial self-healing cement in repairing cracks in cement-based materials and enhancing its resistance to water penetration properties. Cement, a material widely used in infrastructure, has low tensile strength and often forms microcracks. These microcracks reducing the durability of cement and posing risks to the economy and safety. Adding 1% self-healing agent to microbial self-healing cement significantly increases the resistance to water penetration pressure of the mortar blocks. Compared to the standard specimens, the resistance to water penetration ability increased by 29.2% at 28 days and further increased to 39.3% at 84 days. Microbial self-healing cement could effectively restore the resistance to water penetration performance of the mortar blocks after repairing cracks. The repairing results are significantly better than the methods of mixing or applying cement crystalline materials.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 3","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.70010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143521890","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A Cofactor Regeneration System for 2-Aminobutyric Acid Production Based on Combined Cross-Linked Enzyme Aggregates: Utilizing His-Tagged Enzymes With Low-Concentration Calcium Ions as Precipitant 基于复合交联酶聚集体的2-氨基丁酸辅因子再生系统:利用低浓度钙离子的his标记酶作为沉淀剂
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-02-28 DOI: 10.1002/elsc.70013
Jingran Liu, Ren Li, Jincheng Miao, Hongxu Sun, Qiwei Chen, Haiyan Song, Hui Peng, Yanhong Chang, Hui Luo
{"title":"A Cofactor Regeneration System for 2-Aminobutyric Acid Production Based on Combined Cross-Linked Enzyme Aggregates: Utilizing His-Tagged Enzymes With Low-Concentration Calcium Ions as Precipitant","authors":"Jingran Liu,&nbsp;Ren Li,&nbsp;Jincheng Miao,&nbsp;Hongxu Sun,&nbsp;Qiwei Chen,&nbsp;Haiyan Song,&nbsp;Hui Peng,&nbsp;Yanhong Chang,&nbsp;Hui Luo","doi":"10.1002/elsc.70013","DOIUrl":"https://doi.org/10.1002/elsc.70013","url":null,"abstract":"<p>Combined cross-linked enzyme aggregates (combi-CLEAs) represent a promising carrier-free immobilized enzyme technology. This study describes the preparation of combi-CLEAs comprising leucine dehydrogenase (LeuDH) and formate dehydrogenase (FDH) for the regeneration of cofactor nicotinamide adenine dinucleotide necessary for 2-aminobutyric acid production. Different from traditional methods using ammonium sulfate or organic reagents as precipitant, this work utilized low concentrations of calcium ions to purify and precipitate the histidine-tagged enzymes. We developed a simple and environmentally friendly protocol for combi-CLEAs formation, involving precipitation with 10 mM calcium ions at an enzyme activity ratio of 1:2 for LeuDH and FDH, respectively, followed by cross-linking with 0.15% (w/v) glutaraldehyde at 20°C for 2 h at pH 7.5. The optimal catalytic reaction temperature and pH value for the combi-CLEAs were determined to be a temperature of 37°C and a pH of 7.5. The combi-CLEAs demonstrated enhanced thermal and pH tolerance compared to the free enzyme mixture. Moreover, the combi-CLEAs showed good operational stability, retaining 40% of its initial activity after seven cycles of reuse. These findings suggest that the combi-CLEAs of LeuDH and FDH are an efficient and cost-effective option for 2-aminobutyric acid production.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 3","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.70013","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143521767","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
RETRACTION: Evaluation of SDS-Coated Iron Nanostructure on the Gene Expression of Bio Surfactant-Producing Genes by Pseudomonas Aeruginosa 摘要:sds包覆铁纳米结构对铜绿假单胞菌生物表面活性剂产生基因表达的影响
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-02-26 DOI: 10.1002/elsc.70007
{"title":"RETRACTION: Evaluation of SDS-Coated Iron Nanostructure on the Gene Expression of Bio Surfactant-Producing Genes by Pseudomonas Aeruginosa","authors":"","doi":"10.1002/elsc.70007","DOIUrl":"https://doi.org/10.1002/elsc.70007","url":null,"abstract":"<p><b>Retraction</b>: Y.A. Arani, Z. Noormohammadi, B. Rasekh, F. Yazdian, and H. Kazemi, “Evaluation of SDS-Coated Iron Nanostructure on the Gene Expression of Bio Surfactant-Producing Genes by Pseudomonas Aeruginosa,” <i>Engineering in Life Sciences</i> 22, no. 9 (2022): 584–593, https://doi.org/10.1002/elsc.202200002.</p><p>The above article, published online on 24 August 2022, in Wiley Online Library (http://onlinelibrary.wiley.com/), has been retracted by agreement between the journal Editors-in-Chief, An-Ping Zeng and Ralf Takors; and Wiley Periodicals LLC. Following an investigation by the publisher, the parties have concluded that this article was accepted solely on the basis of a compromised peer review process. In addition, a third party informed the publisher that Figures 4 and 5 were reproduced from two articles published either previously or in the same year, and were used here in a different scientific context. The publisher has investigated and confirmed this, and found additional image manipulation in Figure 4. Therefore, the article must be retracted. Corresponding author Behnam Rasekh disagrees with this decision.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 2","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.70007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143497387","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Secretory Production of Heterologous Antimicrobial Peptides in Corynebacterium glutamicum 谷氨酸棒状杆菌分泌产生异源抗菌肽的研究
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-02-18 DOI: 10.1002/elsc.70008
Wei Long, Lina Apitius, Patrick Lenz, Felix Jakob, Anna Joёlle Ruff , Ulrich Schwaneberg
{"title":"Secretory Production of Heterologous Antimicrobial Peptides in Corynebacterium glutamicum","authors":"Wei Long,&nbsp;Lina Apitius,&nbsp;Patrick Lenz,&nbsp;Felix Jakob,&nbsp;Anna Joёlle Ruff ,&nbsp;Ulrich Schwaneberg","doi":"10.1002/elsc.70008","DOIUrl":"https://doi.org/10.1002/elsc.70008","url":null,"abstract":"<p>Antimicrobial peptides (AMPs) are host defense peptides that act against a broad spectrum of microorganisms. AMPs are of high interest as medicinal products, antimicrobial coatings, and for controlling biofilm formation. Applications and research of many AMPs are still hampered by insufficient titers and lack of production platforms that can tolerate high titers of AMPs. <i>Corynebacterium glutamicum</i> is an excellent microbial host for protein secretion and has been barely explored as a host for AMP production. Here, we report the successful production and secretion of two AMPs (amounts of up to 130 mg/L for liquid chromatography peak I [LCI] and 54 mg/L for Psoriasin) by <i>C. glutamicum</i> with low amounts of secreted byproducts.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 2","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.70008","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143438751","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Real-Time VCC Monitoring and Forecasting in HEK-Cell-Based rAAV Vector Production Using Capacitance Spectroscopy 基于hek细胞的rAAV载体生产中VCC的实时监测与预测
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-02-06 DOI: 10.1002/elsc.70004
Rafael Machleid, Suneetha Nunna, Ajith George, Jonas Austerjost, Magda Tomala, Izabella Surowiec
{"title":"Real-Time VCC Monitoring and Forecasting in HEK-Cell-Based rAAV Vector Production Using Capacitance Spectroscopy","authors":"Rafael Machleid,&nbsp;Suneetha Nunna,&nbsp;Ajith George,&nbsp;Jonas Austerjost,&nbsp;Magda Tomala,&nbsp;Izabella Surowiec","doi":"10.1002/elsc.70004","DOIUrl":"https://doi.org/10.1002/elsc.70004","url":null,"abstract":"<p>Recombinant adeno-associated virus (rAAV) vector production is a complex process in which the robust cultivation of human embryonic kidney cells (HEK293) plays a critical role in generating high-quality viral vectors. Tracking the viable cell concentration (VCC) during upstream production is essential for process monitoring and for implementing actions that ensure optimal process management. The advent of inline capacitance probes has introduced a crucial process analytical technology (PAT) tool for real-time VCC measurement. Here, we present the development and application of a method for real-time monitoring of VCC in HEK293-based rAAV vector production. In a first step, BioPAT Viamass probes were used to record capacitance data of individual 10 L rAAV-8 batches within a frequency range of 50 kHz–20 MHz. Based on the capacitance data, a linear single-frequency model and an orthogonal partial least square (OPLS) multifrequency model for VCC prediction were developed. Subsequently, these models were deployed inline, and predictions were exposed into BioPAT MFCS bioprocess control software, enabling real-time VCC monitoring in subsequent rAAV-8 production batches. In addition, the continuous VCC signal was used as input for an exponential cell growth model that was deployed inline to provide accurate real-time forecasting of the transfection time point. To the best of our knowledge, this is the first example of inline deployment of VCC and Time-Till-Transfection predictive models to the bioprocess control system for real-time monitoring and forecasting of these parameters in HEK-cell-based transient rAAV vector production.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 2","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.70004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143362476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Combination of Two-Stage Continuous Feeding and Optimized Synthetic Medium Increases Lipid Production in Lipomyces starkeyi 两段连续饲养与优化合成培养基相结合可提高starkeyi脂菌的产脂量。
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-01-29 DOI: 10.1002/elsc.70003
Chih-Chan Wu, Kenji Okano, Pijar Religia, Yuki Soma, Masatomo Takahashi, Yoshihiro Izumi, Takeshi Bamba, Kohsuke Honda
{"title":"Combination of Two-Stage Continuous Feeding and Optimized Synthetic Medium Increases Lipid Production in Lipomyces starkeyi","authors":"Chih-Chan Wu,&nbsp;Kenji Okano,&nbsp;Pijar Religia,&nbsp;Yuki Soma,&nbsp;Masatomo Takahashi,&nbsp;Yoshihiro Izumi,&nbsp;Takeshi Bamba,&nbsp;Kohsuke Honda","doi":"10.1002/elsc.70003","DOIUrl":"10.1002/elsc.70003","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <p>The oleaginous yeast <i>Lipomyces starkeyi</i> is recognized for its remarkable lipid accumulation under nitrogen-limited conditions. However, precise control of microbial lipid production in <i>L. starkeyi</i> remains challenging due to the complexity of nutrient media.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <p>We developed a two-stage fed-batch fermentation process using a well-defined synthetic medium in a 5-L bioreactor. In the first stage, the specific growth rate was maintained at a designated level by maximizing the cell density through optimizing the feeding rate, molar carbon-to-nitrogen (C/N) ratio, and phosphate concentration in feeding media, achieving a high cell density of 213 ± 10 × 10<sup>7</sup> cells mL<sup>−1</sup>. In the second stage, we optimized the molar C/N ratio in the feeding medium for lipid production and achieved high biomass (130 ± 5 g L<sup>−1</sup>), lipid titer (88 ± 6 g L<sup>−1</sup>), and lipid content (67% ± 2% of dry cellular weight). Our approach yielded a high lipid titer, comparable to the highest reported value of 68 g L<sup>−1</sup> achieved in a nutrient medium, by optimizing cultivation conditions with a synthetic medium in <i>L. starkeyi</i>. This highlights the importance of well-established yet powerful bioprocess approaches for the precise control of microbial cultivation.</p>\u0000 </section>\u0000 </div>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11779743/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143064544","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Integrated SegFlow, µSIA, and UPLC for Online Sialic Acid Quantitation of Glycoproteins Directly from Bioreactors 集成了SegFlow,µSIA和UPLC,用于直接从生物反应器中在线定量糖蛋白唾液酸。
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-01-23 DOI: 10.1002/elsc.202400031
Letha Chemmalil, Tanmay Kulkarni, Mathura Raman, Priya Singh, Yueming Qian, Chris Chumsae, Kyle McHugh, Zhuangrong Huang, Eric Hodgman, Michael C. Borys, Julia Ding, Gloria Li, Anthony Leone
{"title":"Integrated SegFlow, µSIA, and UPLC for Online Sialic Acid Quantitation of Glycoproteins Directly from Bioreactors","authors":"Letha Chemmalil,&nbsp;Tanmay Kulkarni,&nbsp;Mathura Raman,&nbsp;Priya Singh,&nbsp;Yueming Qian,&nbsp;Chris Chumsae,&nbsp;Kyle McHugh,&nbsp;Zhuangrong Huang,&nbsp;Eric Hodgman,&nbsp;Michael C. Borys,&nbsp;Julia Ding,&nbsp;Gloria Li,&nbsp;Anthony Leone","doi":"10.1002/elsc.202400031","DOIUrl":"10.1002/elsc.202400031","url":null,"abstract":"<p>This study emphasizes the critical importance of closely monitoring and controlling the sialic acid content in therapeutic glycoproteins, including EPO, interferon-γ, Orencia, Enbrel, and others, as the level of sialylation directly impacts their pharmacokinetics (PK), immunogenicity, potency, and overall clinical performance due to its influence on protein clearance via hepatic asialoglycoprotein receptors (ASGPR). The ASGPR recognizes and binds to glycoproteins exposed to terminal galactose or N-acetylgalactosamine residues, leading to receptor-mediated endocytosis. Recent studies have demonstrated that sialylation of O-linked glycan plays a role in protecting against macrophage galactose lectin (MGL)-mediated clearance. In addition to the impact on serum half-life, sialylation can influence other clinical outcomes, including immunogenicity, potency, and cytotoxicity. Therefore, the level of sialic acid is a critical quality attribute (CQA), and monitoring and regulating sialylation has become a regulatory requirement to ensure desired clinical performance. To achieve consistent levels of sialic acid-to-protein ratio, the time of upstream harvest and conductivity of downstream wash buffers must be tightly regulated based on the sialic acid content. Therefore, the utilization of process analytical technology (PAT) tools for generating real-time or near-real-time sialic acid content is a business-critical requirement. This work demonstrates the utility of an integrated PAT system for near real-time online sialic acid measurements. The system consists of a micro-sequential injection analyzer (µSIA) interfaced with SegFlow and an ultra performance liquid chromatography (UPLC). The fully automated architecture exemplifies the execution of online sampling, automatic sample preparation, and subsequent online UPLC analysis. This carefully orchestrated PAT framework effectively supports the requirements of QbD-driven continuous bioprocessing.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11756511/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143028216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Evaluation of Yeast Alcohol Acetyltransferases for Ethyl Acetate Production in Clostridium ljungdahlii 永达梭菌生产乙酸乙酯的酵母菌酒精乙酰转移酶的评价
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-01-23 DOI: 10.1002/elsc.202400076
Santiago T. Boto, Kareem Gerges, Bettina Bardl, Miriam A. Rosenbaum
{"title":"Evaluation of Yeast Alcohol Acetyltransferases for Ethyl Acetate Production in Clostridium ljungdahlii","authors":"Santiago T. Boto,&nbsp;Kareem Gerges,&nbsp;Bettina Bardl,&nbsp;Miriam A. Rosenbaum","doi":"10.1002/elsc.202400076","DOIUrl":"10.1002/elsc.202400076","url":null,"abstract":"<p>Sustainable chemical production from C<sub>1</sub> gaseous substrates, such as syngas or CO<sub>2</sub>/H<sub>2</sub>, can be achieved through gas fermentation. In gas fermentation, acetogenic bacteria are able to utilize oxidized inorganic carbon sources as the sole carbon source and electron acceptor, while reduced inorganic species are used as the electron donor. <i>Clostridium ljungdahlii</i>, a model acetogen, is only capable of reducing CO<sub>2</sub> to acetate and ethanol, with H<sub>2</sub> as electron donor. In order to expand the product profile of this bacterium, five alcohol acetyltransferases (AATs) from yeast were heterologously expressed in <i>C. ljungdahlii</i> to evaluate its potential to produce ethyl acetate. When growing on CO<sub>2</sub> and H<sub>2</sub>, up to 7.38 ± 0.43 mg/L of ethyl acetate were produced. Using fructose as the main carbon and energy source, up to 23.15 ± 1.28 mg/L of ethyl acetate were produced. Ethanol and fumarate supplementation were able to boost ethyl acetate titers (up to 37.51 ± 9.44 mg/L). Hence, ethyl acetate production was enabled in <i>C. ljungdahlii</i> at low titers, which could be explained by the high energetic cost of operation of AATs, and their shown promiscuity. However, we also show that this opens the door to more complex esterification reactions of higher added value biomolecules.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11756512/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143028214","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Polyporus umbellatus, A Precious Rare Fungus With Good Pharmaceutical and Food Value 一种具有良好药用和食用价值的珍贵稀有真菌——伞形蓼。
IF 3.9 4区 生物学
Engineering in Life Sciences Pub Date : 2025-01-19 DOI: 10.1002/elsc.202400048
Sizhu Ren, Hua Liu, Qing Sang, Yifan Sun, Liyan Li, Wenjie Chen
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