Engineering in Life Sciences最新文献

{"title":"Cover Picture: Engineering in Life Sciences 11'24","authors":"","doi":"10.1002/elsc.202470111","DOIUrl":"https://doi.org/10.1002/elsc.202470111","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 11","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202470111","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142579724","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}

{"title":"Mechanical Microvibration Device Enhancing Immunohistochemistry Efficiency","authors":"Weifeng Zhang,&nbsp;Jirui Li,&nbsp;Fengshan Xie,&nbsp;Liting Zeng,&nbsp;Liangli Hong,&nbsp;Penghao Li,&nbsp;Xiaomiao Yan,&nbsp;Jingliang Xu,&nbsp;Meina Du,&nbsp;Jiongzhi Hong,&nbsp;Dingrong Yi,&nbsp;Jiahao Xie,&nbsp;Jiang Gu","doi":"10.1002/elsc.202400062","DOIUrl":"https://doi.org/10.1002/elsc.202400062","url":null,"abstract":"<p>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.</p>","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}

{"title":"Cover Picture: Engineering in Life Sciences 10'24","authors":"","doi":"10.1002/elsc.202470101","DOIUrl":"https://doi.org/10.1002/elsc.202470101","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 10","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202470101","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142435271","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}

{"title":"Identification of Plant Peroxidases Catalyzing the Degradation of Fluorinated Aromatics Using a Peroxidase Library Approach","authors":"Ashton Ware,&nbsp;Sally Hess,&nbsp;David Gligor,&nbsp;Sierra Numer,&nbsp;Jack Gregory,&nbsp;Carson Farmer,&nbsp;Gregory M. Raner,&nbsp;Hector E. Medina","doi":"10.1002/elsc.202400054","DOIUrl":"10.1002/elsc.202400054","url":null,"abstract":"<p>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 <i>Cucurbita</i> 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.</p>","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}

{"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 <jats:italic>Pseudomonas taiwanensis</jats:italic> 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 <jats:italic>dnaJ</jats:italic>, RNA polymerase α subunit‐encoding gene <jats:italic>rpoA</jats:italic>, and loss‐of‐function mutations in the <jats:italic>ttgGHI</jats:italic> solvent efflux pump repressor encoded by <jats:italic>ttgV</jats:italic>. 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}

{"title":"Stability of a Mutualistic Escherichia coli Co-Culture During Violacein Production Depends on the Kind of Carbon Source","authors":"Simon Schick,&nbsp;Tobias Müller,&nbsp;Ralf Takors,&nbsp;Georg A. Sprenger","doi":"10.1002/elsc.202400025","DOIUrl":"10.1002/elsc.202400025","url":null,"abstract":"<p>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 <i>Escherichia coli</i> 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.</p>","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}

{"title":"Cover Picture: Engineering in Life Sciences 9'24","authors":"","doi":"10.1002/elsc.202470091","DOIUrl":"https://doi.org/10.1002/elsc.202470091","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 9","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202470091","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142130338","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}

{"title":"Mini Bubble Columns for Miniaturizing Scale‐Down","authors":"Moritz Wild, Ralf Takors","doi":"10.1002/elsc.202400051","DOIUrl":"https://doi.org/10.1002/elsc.202400051","url":null,"abstract":"The successful scale‐up of biotechnological processes from laboratory to industrial scale is crucial for translating innovation to practice. Scale‐down simulators have emerged as indispensable tools in this endeavor, enabling the evaluation of potential hosts’ adaptability to the dynamic conditions encountered in large‐scale fermenters. By simulating these real‐world scenarios, scale‐down simulators facilitate more accurate estimations of host productivity, thereby improving the process of selecting optimal strains for industrial production. Conventional scale‐down systems for detailed intracellular analysis necessitate an elaborate setup comprising interconnected lab‐scale reactors such as stirred tank reactors (STRs) and plug‐flow reactors (PFRs), often proving time‐consuming and resource‐intensive. This work introduces a miniaturized bubble column reactor setup (60 mL working volume), enabling individual and parallel carbon‐limited chemostat fermentations, offering a more efficient and streamlined approach. The industrially relevant organism <jats:italic>Escherichia coli</jats:italic>, chosen as a model organism, is continuously grown and subjected to carbon starvation for 150 s, followed by a return to carbon excess for another 150 s. The cellular response is characterized by the accumulation of the alarmone guanosine pentaphosphate (ppGpp) accompanied by a significant reduction in energy charge, from 0.8 to 0.7, which is rapidly replenished upon reintroduction of carbon availability. Transcriptomic analysis reveals a two‐phase response pattern, with over 200 genes upregulated and downregulated. The initial phase is dominated by the CRP–cAMP‐ and ppGpp‐mediated response to carbon limitation, followed by a shift to stationary phase‐inducing gene expression under the control of stress sigma factors. The system's validity is confirmed through a thorough comparison with a conventional STR/PFR setup. The analysis reveals the potential of the system to effectively reproduce data gathered from conventional STR/PFR setups, showcasing its potential use as a scale‐down simulator integrated in the process of strain development.","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"436 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142218499","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}

{"title":"Microbial Squalene: A Sustainable Alternative for the Cosmetics and Pharmaceutical Industry – A Review","authors":"Saseendran Shalu,&nbsp;Panam Kunnel Raveendranathan Karthikanath,&nbsp;Vinoth Kumar Vaidyanathan,&nbsp;Lars M. Blank,&nbsp;Andrea Germer,&nbsp;Palanisamy Athiyaman Balakumaran","doi":"10.1002/elsc.202400003","DOIUrl":"10.1002/elsc.202400003","url":null,"abstract":"<p>Squalene is a natural triterpenoid and a biosynthetic precursor of steroids and hopanoids in microorganisms, plants, humans, and other animals. Squalene has exceptional properties, such as its antioxidant activity, a high penetrability of the skin, and the ability to trigger the immune system, promoting its application in the cosmetic, sustenance, and pharmaceutical industries. Because sharks are the primary source of squalene, there is a need to identify low-cost, environment friendly, and sustainable alternatives for producing squalene commercially. This shift has prompted scientists to apply biotechnological advances to research microorganisms for synthesizing squalene. This review summarizes recent metabolic and bioprocess engineering strategies in various microorganisms for the biotechnological production of this valuable molecule.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 10","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142218512","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}

{"title":"Cover Picture: Engineering in Life Sciences 8'24","authors":"","doi":"10.1002/elsc.202470081","DOIUrl":"10.1002/elsc.202470081","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 8","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202470081","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141929775","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}