Engineering in Life Sciences最新文献

{"title":"Mini Bubble Columns for Miniaturizing Scale-Down","authors":"Moritz Wild,&nbsp;Ralf Takors","doi":"10.1002/elsc.202400051","DOIUrl":"10.1002/elsc.202400051","url":null,"abstract":"<p>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 <i>Escherichia coli</i>, 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.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 2","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400051","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":"OA","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}

{"title":"Fungal Enzymes for Saccharification of Gamma-Valerolactone-Pretreated White Birch Wood: Optimization of the Production of Talaromyces amestolkiae Cellulolytic Cocktail","authors":"Laura I. de Eugenio,&nbsp;Isabel de la Torre,&nbsp;Felipe de Salas,&nbsp;Francisco Vila,&nbsp;David Alonso,&nbsp;Alicia Prieto,&nbsp;María Jesús Martínez","doi":"10.1002/elsc.202400029","DOIUrl":"10.1002/elsc.202400029","url":null,"abstract":"<p>Lignocellulosic biomass, the most abundant natural resource on earth, can be used for cellulosic ethanol production but requires a pretreatment to improve enzyme access to the polymeric sugars while obtaining value from the other components. γ-Valerolactone (GVL) is a promising candidate for biomass pretreatment since it is renewable and bio-based. In the present work, the effect of a pretreatment based on GVL on the enzymatic saccharification of white birch was evaluated at a laboratory scale and the importance of the washing procedure for the subsequent saccharification was demonstrated. Both the saccharification yield and the production of cellulosic ethanol were higher using a noncommercial enzyme crude from <i>Talaromyces amestolkiae</i> than with the commercial cocktail Cellic CTec2 from Novozymes. Furthermore, the production of extracellular cellulases by <i>T. amestolkiae</i> has been optimized in 2 L bioreactors, with improvements ranging from 40% to 75%. Finally, it was corroborated by isoelectric focus that optimization of cellulase secretion by <i>T. amestolkiae</i> did not affect the pattern production of the main β-glucosidases and endoglucanases secreted by this fungus.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 9","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400029","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141611799","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 7'24","authors":"","doi":"10.1002/elsc.202470071","DOIUrl":"https://doi.org/10.1002/elsc.202470071","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 7","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202470071","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141536958","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":"Improving Downstream Process Related Manufacturability Based on Protein Engineering—A Feasibility Study","authors":"Florian Capito,&nbsp;Ting Hin Wong,&nbsp;Christine Faust,&nbsp;Kilian Brand,&nbsp;Werner Dittrich,&nbsp;Mark Sommerfeld,&nbsp;Garima Tiwari,&nbsp;Thomas Langer","doi":"10.1002/elsc.202400019","DOIUrl":"10.1002/elsc.202400019","url":null,"abstract":"<p>While bioactivity and a favorable safety profile for biotherapeutics is of utmost importance, manufacturability is also worth of consideration to ease the manufacturing process. Manufacturability in the scientific literature is mostly related to stability of formulated drug substances, with limited focus on downstream process-related manufacturability, that is, how easily can a protein be purified. Process-related impurities or biological impurities like viruses and host cell proteins (HCP) are present in the harvest which have mostly acid isoelectric points and need to be removed to ensure patient safety. Therefore, during molecule design, the surface charge of the target molecule should preferably differ sufficiently from the surface charge of the impurities to enable an efficient purification strategy. In this feasibility study, we evaluated the possibility of improving manufacturability by adapting the surface charge of the target protein. We generated several variants of a GLP1-receptor-agonist-Fc-domain-FGF21-fusion protein and demonstrated proof of concept exemplarily for an anion exchange chromatography step which then can be operated at high pH values with maximal product recovery allowing removal of HCP and viruses. Altering the surface charge distribution of biotherapeutic proteins can thus be useful allowing for an efficient manufacturing process for removing HCP and viruses, thereby reducing manufacturing costs.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 9","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400019","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141501456","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":"Editing Streptomyces genome using target AID system fused with UGI-degradation tag","authors":"Pamella Apriliana,&nbsp;Prihardi Kahar,&nbsp;Norimasa Kashiwagi,&nbsp;Akihiko Kondo,&nbsp;Chiaki Ogino","doi":"10.1002/elsc.202400005","DOIUrl":"10.1002/elsc.202400005","url":null,"abstract":"<p>The utilization of <i>Streptomyces</i> as a microbial chassis for developing innovative drugs and medicinal compounds showcases its capability to produce bioactive natural substances. Recent focus on the clustered regularly interspaced short palindromic repeat (CRISPR) technology highlights its potential in genome editing. However, applying CRISPR technology in certain microbial strains, particularly <i>Streptomyces</i>, encounters specific challenges. These challenges include achieving efficient gene expression and maintaining genetic stability, which are critical for successful genome editing. To overcome these obstacles, an innovative approach has been developed that combines several key elements: activation-induced cytidine deaminase (AID), nuclease-deficient cas9 variants (dCas9), and Petromyzon marinus cytidine deaminase 1 (PmCDA1). In this study, this novel strategy was employed to engineer a <i>Streptomyces coelicolor</i> strain. The target gene was actVA-ORF4 (SCO5079), which is involved in actinorhodin production. The engineering process involved introducing a specific construct [pGM1190-dcas9-pmCDA-UGI-AAV-actVA-ORF4 (SCO5079)] to create a CrA10 mutant strain. The resulting CrA10 mutant strain did not produce actinorhodin. This outcome highlights the potential of this combined approach in the genetic manipulation of <i>Streptomyces</i>. The failure of the CrA10 mutant to produce actinorhodin conclusively demonstrates the success of gene editing at the targeted site, affirming the effectiveness of this method for precise genetic modifications in <i>Streptomyces</i>.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 8","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141501502","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 6'24","authors":"","doi":"10.1002/elsc.202470061","DOIUrl":"https://doi.org/10.1002/elsc.202470061","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 6","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202470061","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141251397","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":"Robot-based 6D bioprinting for soft tissue biomedical applications","authors":"Franziska B. Albrecht,&nbsp;Freia F. Schmidt,&nbsp;Christian Schmidt,&nbsp;Rainer Börret,&nbsp;Petra J. Kluger","doi":"10.1002/elsc.202300226","DOIUrl":"10.1002/elsc.202300226","url":null,"abstract":"<p>Within this interdisciplinary study, we demonstrate the applicability of a 6D printer for soft tissue engineering models. For this purpose, a special plant was constructed, combining the technical requirements for 6D printing with the biological necessities, especially for soft tissue. Therefore, a commercial 6D robot arm was combined with a sterilizable housing (including a high-efficiency particulate air (HEPA) filter and ultraviolet radiation (UVC) lamps) and a custom-made printhead and printbed. Both components allow cooling and heating, which is desirable for working with viable cells. In addition, a spraying unit was installed that allows the distribution of fine droplets of a liquid. Advanced geometries on uneven or angled surfaces can be created with the use of all six axes. Based on often used bioinks in the field of soft tissue engineering (gellan gum, collagen, and gelatin methacryloyl) with very different material properties, we could demonstrate the flexibility of the printing system. Furthermore, cell-containing constructs using primary human adipose-derived stem cells (ASCs) could be produced in an automated manner. In addition to cell survival, the ability to differentiate along the adipogenic lineage could also be demonstrated as a representative of soft tissue engineering.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 7","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202300226","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141167669","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":"Genetic modules for α-factor pheromone controlled growth regulation of Saccharomyces cerevisiae","authors":"Uta Gutbier,&nbsp;Juliane Korp,&nbsp;Lennart Scheufler,&nbsp;Kai Ostermann","doi":"10.1002/elsc.202300235","DOIUrl":"10.1002/elsc.202300235","url":null,"abstract":"<p><i>Saccharomyces cerevisiae</i> is a commonly used microorganism in the biotechnological industry. For the industrial heterologous production of compounds, it is of great advantage to work with growth-controllable yeast strains. In our work, we utilized the natural pheromone system of <i>S. cerevisiae</i> and generated a set of different strains possessing an α-pheromone controllable growth behavior. Naturally, the α-factor pheromone is involved in communication between haploid <i>S. cerevisiae</i> cells. Perception of the pheromone initiates several cellular changes, enabling the cells to prepare for an upcoming mating event. We exploited this natural pheromone response system and developed two different plasmid-based modules, in which the target genes, <i>MET15</i> and <i>FAR1</i>, are under control of the α-factor sensitive <i>FIG1</i> promoter for a controlled expression in <i>S. cerevisiae</i>. Whereas expression of <i>MET15</i> led to a growth induction, <i>FAR1</i> expression inhibited growth. The utilization of low copy number or high copy number plasmids for target gene expression and different concentrations of α-factor allow a finely adjustable control of yeast growth rate.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 8","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202300235","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141113331","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}