Laura I. de Eugenio, Isabel de la Torre, Felipe de Salas, Francisco Vila, David Alonso, Alicia Prieto, María Jesús Martínez
{"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, Isabel de la Torre, Felipe de Salas, Francisco Vila, David Alonso, Alicia Prieto, 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":null,"pages":null},"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":null,"pages":null},"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}
Florian Capito, Ting Hin Wong, Christine Faust, Kilian Brand, Werner Dittrich, Mark Sommerfeld, Garima Tiwari, Thomas Langer
{"title":"Improving Downstream Process Related Manufacturability Based on Protein Engineering—A Feasibility Study","authors":"Florian Capito, Ting Hin Wong, Christine Faust, Kilian Brand, Werner Dittrich, Mark Sommerfeld, Garima Tiwari, 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":null,"pages":null},"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, Prihardi Kahar, Norimasa Kashiwagi, Akihiko Kondo, 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":null,"pages":null},"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":null,"pages":null},"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}
Franziska B. Albrecht, Freia F. Schmidt, Christian Schmidt, Rainer Börret, Petra J. Kluger
{"title":"Robot-based 6D bioprinting for soft tissue biomedical applications","authors":"Franziska B. Albrecht, Freia F. Schmidt, Christian Schmidt, Rainer Börret, 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":null,"pages":null},"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}
Uta Gutbier, Juliane Korp, Lennart Scheufler, Kai Ostermann
{"title":"Genetic modules for α-factor pheromone controlled growth regulation of Saccharomyces cerevisiae","authors":"Uta Gutbier, Juliane Korp, Lennart Scheufler, 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":null,"pages":null},"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}
{"title":"Cover Picture: Engineering in Life Sciences 5'24","authors":"","doi":"10.1002/elsc.202470041","DOIUrl":"https://doi.org/10.1002/elsc.202470041","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202470041","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140820487","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}
Feng Ju, Qixiao Zhai, Gang Luo, Hongzhi Tang, Lei Dai
{"title":"Microbiome research for advancing engineering in life science","authors":"Feng Ju, Qixiao Zhai, Gang Luo, Hongzhi Tang, Lei Dai","doi":"10.1002/elsc.202400028","DOIUrl":"10.1002/elsc.202400028","url":null,"abstract":"<p>Microbiome research has become increasingly prominent, as scientists explore the intricately assembled microbial communities (i.e., microbiota) and their wide-ranging impacts on human systems (e.g., health and foods), environmental sustainability (bioremediation, biogeochemistry, and ecosystem biorestoration, or 3B for Sustainability), and next-generation bioeconomy (i.e., bioenergy, biomedicine, and biomaterials, or 3B for Resources). This burgeoning field has been driven by the widespread adoption of meta-omics methodologies, such as metagenomics, metatranscriptomics, metaproteomics, and metabolomics. In this special issue, we present a compendium of recent human and environmental microbiome studies that elucidate the multifaceted roles of microbial communities and their implications across different domains of research in life sciences and related fields of application.</p><p>The gut microbiome stands out as a central player in human health, influencing fundamental physiological processes such as digestion, immunity, and metabolism. Tang et al. delves into the intricate interplay between the gut microbiota and the host epigenome in the context of Non-alcoholic Fatty Liver Disease (NAFLD), shedding light on how microbial factors can modulate gene expression patterns associated with NAFLD pathogenesis [<span>1</span>]. Similarly, Zoghi et al. investigate the association between gut dysbiosis and nutritional imbalances in children, underscoring the potential therapeutic avenues for modulating gut microbiota composition to restore energy homeostasis [<span>2</span>].</p><p>Moreover, the symbiotic interplay between flavonoids and the gut microbiota emerges as a promising area of study in maintaining metabolic balance and overall health by Zhou et al. [<span>3</span>]. Flavonoids, abundant in fruits and vegetables, serve as essential dietary components that undergo biotransformation by gut microbes, yielding bioactive metabolites with various health-promoting properties. Understanding this intricate interplay opens new avenues for leveraging dietary interventions to modulate gut microbiota composition and enhance metabolic health (Figure 1).</p><p>Beyond human health, microbial communities also play critical roles in environmental processes, particularly in the biodegradation of pollutants. Huang et al. leverage meta-omics approaches to uncover the genetic potential of microbial communities in contaminated environments, offering insights into potential bioremediation strategies for mitigating environmental pollution [<span>4</span>].</p><p>Furthermore, microbiomes offer promising avenues for bioconversion and biodegradation processes in the context of biotechnology and industrial applications. Zhu et al. investigate the dynamics of microbial consortia during primary sludge and food waste fermentation, revealing insights into how different environmental conditions and additives can modulate fermentation product profiles [<span>5</span>]. The study","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400028","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140566527","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 4'24","authors":"","doi":"10.1002/elsc.202470033","DOIUrl":"https://doi.org/10.1002/elsc.202470033","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202470033","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140345747","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}