Engineering in Life Sciences最新文献

Cover Picture: Engineering in Life Sciences 12'24

IF 3.9 4区生物学

Engineering in Life Sciences Pub Date : 2024-12-05 DOI: 10.1002/elsc.202470121

引用次数: 0

Cover Picture: Engineering in Life Sciences 11'24 封面图片：生命科学工程 11'24

IF 3.9 4区生物学

Engineering in Life Sciences Pub Date : 2024-11-04 DOI: 10.1002/elsc.202470111

引用次数: 0

Raman-Enabled Predictions of Protein Content and Metabolites in Biopharmaceutical Saccharomyces cerevisiae Fermentations

IF 3.9 4区生物学

Engineering in Life Sciences Pub Date : 2024-10-16 DOI: 10.1002/elsc.202400045

Jeppe Hagedorn, Guilherme Ramos, Miguel Ressurreição, Ernst Broberg Hansen, Michael Sokolov, Carlos Casado Vázquez, Christos Panos

{"title":"Raman-Enabled Predictions of Protein Content and Metabolites in Biopharmaceutical Saccharomyces cerevisiae Fermentations","authors":"Jeppe Hagedorn, Guilherme Ramos, Miguel Ressurreição, Ernst Broberg Hansen, Michael Sokolov, Carlos Casado Vázquez, Christos Panos","doi":"10.1002/elsc.202400045","DOIUrl":"10.1002/elsc.202400045","url":null,"abstract":"Raman spectroscopy, a robust and non-invasive analytical method, has demonstrated significant potential for monitoring biopharmaceutical production processes. Its ability to provide detailed information about molecular vibrations makes it ideal for the detection and quantification of therapeutic proteins and critical control parameters in complex biopharmaceutical mixtures. However, its application in Saccharomyces cerevisiae fermentations has been hindered by the inherent strong fluorescence background from the cells. This fluorescence interferes with Raman signals, compromising spectral data accuracy. In this study, we present an approach that mitigates this issue by deploying Raman spectroscopy on cell-free media samples, combined with advanced chemometric modeling. This method enables accurate prediction of protein concentration and key process parameters, fundamental for the control and optimization of biopharmaceutical fermentation processes. Utilizing variable importance in projection (VIP) further enhances model robustness, leading to lower relative root mean squared error of prediction (RMSEP) values across the six targets studied. Our findings highlight the potential of Raman spectroscopy for real-time, on-line monitoring and control of complex microbial fermentations, thereby significantly enhancing the efficiency and quality of S. cerevisiae-based biopharmaceutical production.","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 12","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11620617/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142794628","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

Mechanical Microvibration Device Enhancing Immunohistochemistry Efficiency 提高免疫组化效率的机械微振动装置

IF 3.9 4区生物学

Engineering in Life Sciences Pub Date : 2024-10-13 DOI: 10.1002/elsc.202400062

Weifeng Zhang, Jirui Li, Fengshan Xie, Liting Zeng, Liangli Hong, Penghao Li, Xiaomiao Yan, Jingliang Xu, Meina Du, Jiongzhi Hong, Dingrong Yi, Jiahao Xie, Jiang Gu

{"title":"Mechanical Microvibration Device Enhancing Immunohistochemistry Efficiency","authors":"Weifeng Zhang, Jirui Li, Fengshan Xie, Liting Zeng, Liangli Hong, Penghao Li, Xiaomiao Yan, Jingliang Xu, Meina Du, Jiongzhi Hong, Dingrong Yi, Jiahao Xie, Jiang Gu","doi":"10.1002/elsc.202400062","DOIUrl":"https://doi.org/10.1002/elsc.202400062","url":null,"abstract":"Immunohistochemistry (IHC) is a widely used technique in diagnostic pathology and biomedical research, but there is still a need to shorten the operation process and reduce the cost of antibodies. This study aims to assess a novel IHC technique that incorporates mechanical microvibration (MMV) to expedite the process, reduce antibody consumption, and enhance staining quality. MMV was generated using coin vibration motors attached to glass slides mounted with consecutive tissue sections. Multiple antibodies targeting various antigens were used to stain cancerous and normal tissues, with and without microvibration. Various parameters were tested, including incubation durations, temperatures, and antibody dilutions. The novel method showed the potential to achieve comparable or superior outcomes in significantly less time, utilizing over 10 times less antibody than controls. MMV improved specific staining quality, yielding stronger, and better-defined positive reactions. This was validated through a multicenter double-blind assessment and quantitative image analysis. The possible mechanisms were also investigated. MMV shortens immunohistochemical staining duration, reduces antibody usage, and enhances staining specificity, likely by accelerating antibody movement and diffusion. These improvements translate to time and cost savings, offering clinical and financial value for diagnostic pathology and biomedical research.","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 11","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400062","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142579762","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

Cover Picture: Engineering in Life Sciences 10'24 封面图片：生命科学工程 10'24

IF 3.9 4区生物学

Engineering in Life Sciences Pub Date : 2024-10-09 DOI: 10.1002/elsc.202470101

引用次数: 0

In situ Product Recovery of Microbially Synthesized Ethyl Acetate from the Exhaust Gas of a Bioreactor by Membrane Technology

IF 3.9 4区生物学

Engineering in Life Sciences Pub Date : 2024-09-30 DOI: 10.1002/elsc.202400041

Andreas Hoffmann, Alexander Franz, Christian Löser, Thomas Hoyer, Marcus Weyd, Thomas Walther

{"title":"In situ Product Recovery of Microbially Synthesized Ethyl Acetate from the Exhaust Gas of a Bioreactor by Membrane Technology","authors":"Andreas Hoffmann, Alexander Franz, Christian Löser, Thomas Hoyer, Marcus Weyd, Thomas Walther","doi":"10.1002/elsc.202400041","DOIUrl":"10.1002/elsc.202400041","url":null,"abstract":"Ethyl acetate is at present exclusively produced from fossil resources. Microbial synthesis of this ester from sugar-rich waste as an alternative is an aerobic process. Ethyl acetate is highly volatile and therefore stripped with the exhaust gas from the bioreactor which enables in situ product recovery. Previous research on microbial formation of ethyl acetate has focused on the kinetics of ester synthesis and in part on the ester stripping, while the separation of the ester from the exhaust gas has hardly been investigated. A mixed matrix membrane was developed consisting of Silikalite-1 embedded in polydimethylsiloxane which was installed in a radial–symmetrical membrane module. Evaluation of the separation of ethyl acetate was based on the analysis of the composition of the feed and retentate gas by mass spectrometry. The separation efficiency of the membrane was first tested with varied flows of artificial exhaust gas, containing defined amounts of ethyl acetate. A model for describing the separation process was parametrized by the measured data and used to design a real separation experiment. Ethyl acetate produced from delactosed whey permeate by Kluyveromyces marxianus DSM 5422 in a stirred bioreactor gassed with 0.5 vvm air was successfully separated from the exhaust gas by membranes; 93.6% of the stripped ester was separated. Liquid ethyl acetate was recovered by cooling the permeate gas to ‒78°C, whereby 99.75% of the condensed organic compounds were ethyl acetate. This study demonstrates for the first time that microbially produced and stripped ethyl acetate can be effectively separated from the exhaust gas of bioreactors by membrane technology to obtain the ester in high yield and purity.","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 12","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11620624/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142794626","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

Identification of Plant Peroxidases Catalyzing the Degradation of Fluorinated Aromatics Using a Peroxidase Library Approach 利用过氧化物酶库方法鉴定催化氟化芳烃降解的植物过氧化物酶

IF 3.9 4区生物学

Engineering in Life Sciences Pub Date : 2024-09-17 DOI: 10.1002/elsc.202400054

Ashton Ware, Sally Hess, David Gligor, Sierra Numer, Jack Gregory, Carson Farmer, Gregory M. Raner, Hector E. Medina

{"title":"Identification of Plant Peroxidases Catalyzing the Degradation of Fluorinated Aromatics Using a Peroxidase Library Approach","authors":"Ashton Ware, Sally Hess, David Gligor, Sierra Numer, Jack Gregory, Carson Farmer, Gregory M. Raner, Hector E. Medina","doi":"10.1002/elsc.202400054","DOIUrl":"10.1002/elsc.202400054","url":null,"abstract":"In this work, the degradation of mono- and polyfluorinated phenolic compounds was demonstrated by a series of crude plant peroxidases, including horseradish root (HRP) and six members of the Cucurbita genus. Highly active samples were identified using a library screening approach in which more than 50 crude plant samples were initially evaluated for defluorination activity toward 4-fluorophenol. The highest concentrations were observed in the HRP, pumpkin skin (PKS), and butternut squash skin (BNS), which consistently gave the highest intrinsic rates of decomposition for all the substrates tested. Although HRP exhibited a significant decrease in activity with increased fluorination of the phenolic substrate, PKS showed only minor reductions. Furthermore, in silico studies indicated that the active site of HRP poorly accommodates the steric bulk of additional fluorines, causing the substrate to dock farther from the catalytic heme and thus slowing the catalysis rate. We propose that the PKS active site might be larger, allowing closer access to the perfluorinated substrate, and therefore maintaining higher activity compared to the HRP enzyme. However, detailed kinetic characterization studies of the peroxidases are recommended. Conclusively, the high catalytic activity of PKS and its high yield per gram of tissue make it an excellent candidate for developing environmentally friendly biocatalytic methods for degrading fluorinated aromatics. Finally, the success of the library approach in identifying highly active samples for polyfluorinated aromatic compound (PFAC) degradation suggests the method may find utility in the quest for other advanced catalysts for PFAS degradation.","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 11","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400054","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142258823","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

SIGHT—A System for Solvent‐Tight Incubation and Growth Monitoring in High Throughput SIGHT--用于高通量溶剂密闭培养和生长监控的系统

IF 2.7 4区生物学

Engineering in Life Sciences Pub Date : 2024-09-11 DOI: 10.1002/elsc.202400037

Jakob Rönitz, Felix Herrmann, Benedikt Wynands, Tino Polen, Nick Wierckx

{"title":"SIGHT—A System for Solvent‐Tight Incubation and Growth Monitoring in High Throughput","authors":"Jakob Rönitz, Felix Herrmann, Benedikt Wynands, Tino Polen, Nick Wierckx","doi":"10.1002/elsc.202400037","DOIUrl":"https://doi.org/10.1002/elsc.202400037","url":null,"abstract":"Physiological characterization of microorganisms in the context of solvent tolerance is a tedious process with a high investment of manual labor while often being limited in throughput capability simultaneously. Therefore, we developed a small‐scale solvent‐impervious cultivation system consisting of screw cap‐sealed glass vials in combination with a 3D‐printed vial holder for the Growth Profiler (EnzyScreen) platform. Components and cultivation conditions were empirically tested, and a suitable setup was found for the intended application. To demonstrate the capability of this cultivation system, an adaptive laboratory evolution was performed to further increase the tolerance of Pseudomonas taiwanensis GRC3 toward styrene. This approach yielded heterogenic cultures with improved growth performances in the presence of styrene from which individual clones were isolated and characterized in high throughput. Several clones with improved growth in the presence of 1% (v/v) styrene were analyzed through whole‐genome sequencing, revealing mutations in the co‐chaperone‐encoding gene dnaJ, RNA polymerase α subunit‐encoding gene rpoA, and loss‐of‐function mutations in the ttgGHI solvent efflux pump repressor encoded by ttgV. The developed cultivation system has proven to be a very useful extension of the Growth Profiler, as it reduces manual workload and allows high‐throughput characterization.","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"37 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142218482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Stability of a Mutualistic Escherichia coli Co-Culture During Violacein Production Depends on the Kind of Carbon Source 互助型大肠杆菌共培养菌群在生产维拉丝素过程中的稳定性取决于碳源种类

IF 3.9 4区生物学

Engineering in Life Sciences Pub Date : 2024-09-08 DOI: 10.1002/elsc.202400025

Simon Schick, Tobias Müller, Ralf Takors, Georg A. Sprenger

{"title":"Stability of a Mutualistic Escherichia coli Co-Culture During Violacein Production Depends on the Kind of Carbon Source","authors":"Simon Schick, Tobias Müller, Ralf Takors, Georg A. Sprenger","doi":"10.1002/elsc.202400025","DOIUrl":"10.1002/elsc.202400025","url":null,"abstract":"The L-tryptophan–derived purple pigment violacein (VIO) is produced in recombinant bacteria and studied for its versatile applications. Microbial synthetic co-cultures are gaining more importance as efficient factories for synthesizing high-value compounds. In this work, a mutualistic and cross-feeding Escherichia coli co-culture is metabolically engineered to produce VIO. The strains are genetically modified by auxotrophies in the tryptophan (TRP) pathway to enable a metabolic division of labor. Therein, one strain produces anthranilate (ANT) and the other transforms it into TRP and further to VIO. Population dynamics and stability depend on the choice of carbon source, impacting the presence and thus exchange of metabolites as well as overall VIO productivity. Four carbon sources (D-glucose, glycerol, D-galactose, and D-xylose) were compared. D-Xylose led to co-cultures which showed stable growth and VIO production, ANT-TRP exchange, and enhanced VIO production. Best titers were ∼126 mg L–1 in shake flasks. The study demonstrates the importance and advantages of a mutualistic approach in VIO synthesis and highlights the carbon source's role in co-culture stability and productivity. Transferring this knowledge into an up-scaled bioreactor system has great potential in improving the overall VIO production.","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 10","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400025","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142218481","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

Cover Picture: Engineering in Life Sciences 9'24 封面图片：生命科学工程 9'24

IF 3.9 4区生物学

Engineering in Life Sciences Pub Date : 2024-09-03 DOI: 10.1002/elsc.202470091

引用次数: 0