Synthetic biology (Oxford, England)最新文献

{"title":"Promoter engineering for microbial bio-alkane gas production.","authors":"Duangthip Trisrivirat,&nbsp;John M X Hughes,&nbsp;Robin Hoeven,&nbsp;Matthew Faulkner,&nbsp;Helen Toogood,&nbsp;Pimchai Chaiyen,&nbsp;Nigel S Scrutton","doi":"10.1093/synbio/ysaa022","DOIUrl":"https://doi.org/10.1093/synbio/ysaa022","url":null,"abstract":"<p><p>Successful industrial biotechnological solutions to biofuels and other chemicals production rely on effective competition with existing lower-cost natural sources and synthetic chemistry approaches enabled by adopting low-cost bioreactors and processes. This is achievable by mobilizing <i>Halomonas</i> as a next generation industrial chassis, which can be cultivated under non-sterile conditions. To increase the cost effectiveness of an existing sustainable low carbon bio-propane production strategy, we designed and screened a constitutive promoter library based on the known strong porin promoter from <i>Halomonas</i>. Comparative studies were performed between <i>Escherichia coli</i> and <i>Halomonas</i> using the reporter gene red fluorescent protein (RFP). Later studies with a fatty acid photodecarboxylase-RFP fusion protein demonstrated tuneable propane production in <i>Halomonas</i> and <i>E. coli</i>, with an ∼8-fold improvement in yield over comparable isopropyl-β-D-thiogalactoside-inducible systems. This novel set of promoters is a useful addition to the synthetic biology toolbox for future engineering of <i>Halomonas</i> to make chemicals and fuels.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa022"},"PeriodicalIF":0.0,"publicationDate":"2020-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysaa022","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38324123","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advances in engineering CRISPR-Cas9 as a molecular Swiss Army knife.","authors":"Grace A Meaker, Emma J Hair, Thomas E Gorochowski","doi":"10.1093/synbio/ysaa021","DOIUrl":"10.1093/synbio/ysaa021","url":null,"abstract":"<p><p>The RNA-guided endonuclease system CRISPR-Cas9 has been extensively modified since its discovery, allowing its capabilities to extend far beyond double-stranded cleavage to high fidelity insertions, deletions and single base edits. Such innovations have been possible due to the modular architecture of CRISPR-Cas9 and the robustness of its component parts to modifications and the fusion of new functional elements. Here, we review the broad toolkit of CRISPR-Cas9-based systems now available for diverse genome-editing tasks. We provide an overview of their core molecular structure and mechanism and distil the design principles used to engineer their diverse functionalities. We end by looking beyond the biochemistry and toward the societal and ethical challenges that these CRISPR-Cas9 systems face if their transformative capabilities are to be deployed in a safe and acceptable manner.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa021"},"PeriodicalIF":0.0,"publicationDate":"2020-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysaa021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38735379","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>In silico</i> design and automated learning to boost next-generation smart biomanufacturing.","authors":"Pablo Carbonell,&nbsp;Rosalind Le Feuvre,&nbsp;Eriko Takano,&nbsp;Nigel S Scrutton","doi":"10.1093/synbio/ysaa020","DOIUrl":"https://doi.org/10.1093/synbio/ysaa020","url":null,"abstract":"<p><p>The increasing demand for bio-based compounds produced from waste or sustainable sources is driving biofoundries to deliver a new generation of prototyping biomanufacturing platforms. Integration and automation of the design, build, test and learn (DBTL) steps in centers like SYNBIOCHEM in Manchester and across the globe (Global Biofoundries Alliance) are helping to reduce the delivery time from initial strain screening and prototyping towards industrial production. Notably, a portfolio of producer strains for a suite of material monomers was recently developed, some approaching industrial titers, in a <i>tour de force</i> by the Manchester Centre that was achieved in less than 90 days. New <i>in silico</i> design tools are providing significant contributions to the front end of the DBTL pipelines. At the same time, the far-reaching initiatives of modern biofoundries are generating a large amount of high-dimensional data and knowledge that can be integrated through automated learning to expedite the DBTL cycle. In this Perspective, the new design tools and the role of the learning component as an enabling technology for the next generation of automated biofoundries are discussed. Future biofoundries will operate under completely automated DBTL cycles driven by <i>in silico</i> optimal experimental planning, full biomanufacturing devices connectivity, virtualization platforms and cloud-based design. The automated generation of robotic build worklists and the integration of machine-learning algorithms will collectively allow high levels of adaptability and rapid design changes toward fully automated smart biomanufacturing.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa020"},"PeriodicalIF":0.0,"publicationDate":"2020-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysaa020","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38735377","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modular cell-free expression plasmids to accelerate biological design in cells.","authors":"Ashty S Karim, Fungmin Eric Liew, Shivani Garg, Bastian Vögeli, Blake J Rasor, Aislinn Gonnot, Marilene Pavan, Alex Juminaga, Séan D Simpson, Michael Köpke, Michael C Jewett","doi":"10.1093/synbio/ysaa019","DOIUrl":"10.1093/synbio/ysaa019","url":null,"abstract":"<p><p>Industrial biotechnology aims to produce high-value products from renewable resources. This can be challenging because model microorganisms-organisms that are easy to use like <i>Escherichia coli</i>-often lack the machinery required to utilize desired feedstocks like lignocellulosic biomass or syngas. Non-model organisms, such as <i>Clostridium</i>, are industrially proven and have desirable metabolic features but have several hurdles to mainstream use. Namely, these species grow more slowly than conventional laboratory microbes, and genetic tools for engineering them are far less prevalent. To address these hurdles for accelerating cellular design, cell-free synthetic biology has matured as an approach for characterizing non-model organisms and rapidly testing metabolic pathways <i>in vitro</i>. Unfortunately, cell-free systems can require specialized DNA architectures with minimal regulation that are not compatible with cellular expression. In this work, we develop a modular vector system that allows for T7 expression of desired enzymes for cell-free expression and direct Golden Gate assembly into <i>Clostridium</i> expression vectors. Utilizing the Joint Genome Institute's DNA Synthesis Community Science Program, we designed and synthesized these plasmids and genes required for our projects allowing us to shuttle DNA easily between our <i>in vitro</i> and <i>in vivo</i> experiments. We next validated that these vectors were sufficient for cell-free expression of functional enzymes, performing on par with the previous state-of-the-art. Lastly, we demonstrated automated six-part DNA assemblies for <i>Clostridium autoethanogenum</i> expression with efficiencies ranging from 68% to 90%. We anticipate this system of plasmids will enable a framework for facile testing of biosynthetic pathways <i>in vitro</i> and <i>in vivo</i> by shortening development cycles.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa019"},"PeriodicalIF":0.0,"publicationDate":"2020-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/7f/cc/ysaa019.PMC7737004.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38735376","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Construction of an artificial biosynthetic pathway for hyperextended archaeal membrane lipids in the bacterium <i>Escherichia coli</i>.","authors":"Ryo Yoshida,&nbsp;Hisashi Hemmi","doi":"10.1093/synbio/ysaa018","DOIUrl":"https://doi.org/10.1093/synbio/ysaa018","url":null,"abstract":"<p><p>Archaea produce unique membrane lipids, which possess two fully saturated isoprenoid chains linked to the glycerol moiety via ether bonds. The isoprenoid chain length of archaeal membrane lipids is believed to be important for some archaea to thrive in extreme environments because the hyperthermophilic archaeon <i>Aeropyrum pernix</i> and some halophilic archaea synthesize extended C25,C25-archaeal diether-type membrane lipids, which have isoprenoid chains that are longer than those of typical C20,C20-diether lipids. Natural archaeal diether lipids possessing longer C30 or C35 isoprenoid chains, however, have yet to be isolated. In the present study, we attempted to synthesize such hyperextended archaeal membrane lipids. We investigated the substrate preference of the enzyme <i>sn</i>-2,3-(digeranylfarnesyl)glycerol-1-phosphate synthase from <i>A. pernix</i>, which catalyzes the transfer of the second C25 isoprenoid chain to the glycerol moiety in the biosynthetic pathway of C25,C25-archaeal membrane lipids. The enzyme was shown to accept <i>sn</i>-3-hexaprenylglycerol-1-phosphate, which has a C30 isoprenoid chain, as a prenyl acceptor substrate to synthesize <i>sn</i>-2-geranylfarnesyl-3-hexaprenylglycerol-1-phosphate, a supposed precursor for hyperextended C25,C30-archaeal membrane lipids. Furthermore, we constructed an artificial biosynthetic pathway by introducing 4 archaeal genes and 1 gene from <i>Bacillus subtilis</i> in the cells of <i>Escherichia coli</i>, which enabled the <i>E. coli</i> strain to produce hyperextended C25,C30-archaeal membrane lipids, which have never been reported so far.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa018"},"PeriodicalIF":0.0,"publicationDate":"2020-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysaa018","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38324122","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Elucidation and refinement of synthetic receptor mechanisms.","authors":"Hailey I Edelstein, Patrick S Donahue, Joseph J Muldoon, Anthony K Kang, Taylor B Dolberg, Lauren M Battaglia, Everett R Allchin, Mihe Hong, Joshua N Leonard","doi":"10.1093/synbio/ysaa017","DOIUrl":"10.1093/synbio/ysaa017","url":null,"abstract":"<p><p>Synthetic receptors are powerful tools for engineering mammalian cell-based devices. These biosensors enable cell-based therapies to perform complex tasks such as regulating therapeutic gene expression in response to sensing physiological cues. Although multiple synthetic receptor systems now exist, many aspects of receptor performance are poorly understood. In general, it would be useful to understand how receptor design choices influence performance characteristics. In this study, we examined the modular extracellular sensor architecture (MESA) and systematically evaluated previously unexamined design choices, yielding substantially improved receptors. A key finding that might extend to other receptor systems is that the choice of transmembrane domain (TMD) is important for generating high-performing receptors. To provide mechanistic insights, we adopted and employed a Förster resonance energy transfer-based assay to elucidate how TMDs affect receptor complex formation and connected these observations to functional performance. To build further insight into these phenomena, we developed a library of new MESA receptors that sense an expanded set of ligands. Based upon these explorations, we conclude that TMDs affect signaling primarily by modulating intracellular domain geometry. Finally, to guide the design of future receptors, we propose general principles for linking design choices to biophysical mechanisms and performance characteristics.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa017"},"PeriodicalIF":0.0,"publicationDate":"2020-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7759213/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38776012","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CRAGE-mediated insertion of fluorescent chromosomal markers for accurate and scalable measurement of co-culture dynamics in <i>Escherichia coli</i>.","authors":"Avery J C Noonan,&nbsp;Yilin Qiu,&nbsp;Joe C H Ho,&nbsp;Jewel Ocampo,&nbsp;K A Vreugdenhil,&nbsp;R Alexander Marr,&nbsp;Zhiying Zhao,&nbsp;Yasuo Yoshikuni,&nbsp;Steven J Hallam","doi":"10.1093/synbio/ysaa015","DOIUrl":"https://doi.org/10.1093/synbio/ysaa015","url":null,"abstract":"<p><p>Monitoring population dynamics in co-culture is necessary in engineering microbial consortia involved in distributed metabolic processes or biosensing applications. However, it remains difficult to measure strain-specific growth dynamics in high-throughput formats. This is especially vexing in plate-based functional screens leveraging whole-cell biosensors to detect specific metabolic signals. Here, we develop an experimental high-throughput co-culture system to measure and model the relationship between fluorescence and cell abundance, combining chassis-independent recombinase-assisted genome engineering (CRAGE) and whole-cell biosensing with a P_emrR-green fluorescent protein (GFP) monoaromatic reporter used in plate-based functional screening. CRAGE was used to construct <i>Escherichia coli</i> EPI300 strains constitutively expressing red fluorescent protein (RFP) and the relationship between RFP expression and optical density (OD₆₀₀) was determined throughout the EPI300 growth cycle. A linear equation describing the increase of normalized RFP fluorescence during deceleration phase was derived and used to predict biosensor strain dynamics in co-culture. Measured and predicted values were compared using flow cytometric detection methods. Induction of the biosensor lead to increased GFP fluorescence normalized to biosensor cell abundance, as expected, but a significant decrease in relative abundance of the biosensor strain in co-culture and a decrease in bulk GFP fluorescence. Taken together, these results highlight sensitivity of population dynamics to variations in metabolic activity in co-culture and the potential effect of these dynamics on the performance of functional screens in plate-based formats. The engineered strains and model used to evaluate these dynamics provide a framework for optimizing growth of synthetic co-cultures used in screening, testing and pathway engineering applications.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa015"},"PeriodicalIF":0.0,"publicationDate":"2020-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysaa015","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38768150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rosa26 docking sites for investigating genetic circuit silencing in stem cells.","authors":"Michael Fitzgerald, Mark Livingston, Chelsea Gibbs, Tara L Deans","doi":"10.1093/synbio/ysaa014","DOIUrl":"10.1093/synbio/ysaa014","url":null,"abstract":"<p><p>Approaches in mammalian synthetic biology have transformed how cells can be programmed to have reliable and predictable behavior, however, the majority of mammalian synthetic biology has been accomplished using immortalized cell lines that are easy to grow and easy to transfect. Genetic circuits that integrate into the genome of these immortalized cell lines remain functional for many generations, often for the lifetime of the cells, yet when genetic circuits are integrated into the genome of stem cells gene silencing is observed within a few generations. To investigate the reactivation of silenced genetic circuits in stem cells, the Rosa26 locus of mouse pluripotent stem cells was modified to contain docking sites for site-specific integration of genetic circuits. We show that the silencing of genetic circuits can be reversed with the addition of sodium butyrate, a histone deacetylase inhibitor. These findings demonstrate an approach to reactivate the function of genetic circuits in pluripotent stem cells to ensure robust function over many generations. Altogether, this work introduces an approach to overcome the silencing of genetic circuits in pluripotent stem cells that may enable the use of genetic circuits in pluripotent stem cells for long-term function.</p>","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa014"},"PeriodicalIF":0.0,"publicationDate":"2020-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/67/56/ysaa014.PMC7644442.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38615937","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Designing for durability: new tools to build stable, non-repetitive DNA.","authors":"Pablo Cárdenas","doi":"10.1093/synbio/ysaa016","DOIUrl":"https://doi.org/10.1093/synbio/ysaa016","url":null,"abstract":"The survival of genetic information hinges on identifying repetition. Genomes are repaired by mechanisms such as homologous recombination, in which matching DNA sequences are used as a template to replace missing information. This strategy works provided sequences in the genome are mostly unique. While sequence diversity has kept genomes stable enough to replicate for millions of years, it poses a problem for those trying to engineer DNA (1). After all, one of the central tenets of synthetic biology is the reutilization of standard parts. How, then, can we design stable, non-repetitive genetic systems with a limited toolkit of synthetic parts? Researchers in Howard Salis’s lab at Pennsylvania State University set out to address this challenge through the Non-Repetitive Parts Calculator (NRPC), a set of new algorithms described in a recent publication by Hossain et al. (2) and available online (https://sali slab.net/software/). As the name implies, NRPC builds collections of biological parts containing minimal repetitive sequences, where the repetitiveness of a collection is defined by Lmax, the maximum length of the longest shared repeat. Collections can be created using two different modes. The ‘Finder’ mode determines the largest subset of nonrepetitive elements within any given database of parts, given a maximum Lmax set by the user. The sheer number of possible subsets to evaluate can make this computationally impractical for large libraries. The authors solve this problem by representing parts as nodes on a graph and improving on existing algorithms in graph theory to efficiently maximize the number of disconnected components. The ‘Maker’ mode creates a new library of non-repetitive parts within the design constraints set by the user, which may include a degenerate DNA sequence or RNA structure template and a set Lmax value. In this case, all possible sequences are represented as a decision tree and hash tables are used to store and check for occurrences of sub-sequences within parts. Hossain et al. tested their new ‘Maker’ algorithm by generating libraries of 4350 synthetic, non-repetitive bacterial promoters and 1722 yeast promoters, designed to have a wide range of transcription rates. The authors validated each library’s predicted transcriptional behavior by assembling and characterizing every promoter through next-generation DNA and RNA sequencing in Escherichia coli and Saccharomyces cerevisiae. The increased stability of NRPC designs was demonstrated in E. coli by comparing versions of a construct with either repetitive or non-repetitive promoters. The former rapidly lost fluorescence and DNA content while the latter remained stable. Finally, the authors applied regression models and neural networks developed elsewhere (3) to explain and predict the strength of the synthetic promoters they created. This work can have tremendous, immediate impact in two ways. Not only did Hossain et al. produce vast libraries of bacterial and yeast pro","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa016"},"PeriodicalIF":0.0,"publicationDate":"2020-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysaa016","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38624017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reprogramming the proteome for cell-free protein expression.","authors":"Tetsuhiro Harimoto","doi":"10.1093/synbio/ysaa013","DOIUrl":"https://doi.org/10.1093/synbio/ysaa013","url":null,"abstract":"Living cells produce countless numbers of valuable compounds. By engineering these living ‘biofactories’, synthetic biologists have been making novel molecules that can be used for medicine, food, energy and everyday applications. However, crosstalk between engineered modules and host factors can significantly interfere with biomolecule production by competing for common resources. To address this challenge, scientists have been trying to minimize unwanted crosstalk between the host and synthetic networks by deleting proteins that drain resources such as proteases and metabolizing enzymes. In a recent study in the journal Nature Communications, the research team from Cheemeng Tan’s group at the University of California in Davis took a novel approach (1). Instead of minimizing crosstalk, they intentionally re-engineered crosstalk between the host and synthetic networks in order to create a more favorable environment for protein synthesis. This ‘holistic’ engineering approach achieved a global proteome reprogramming and enabled the production of complex proteins. The research team, led by Luis E. Contreras-Llano and Conary Meyer, utilized a cell-free protein synthesis system to construct consortia of bacteria, each expressing core proteins involved with protein translation. Because expressing multiple proteins in a single strain results in high metabolic burdens to the cells, distribution of the labor between the members of the consortium can improve overall protein expression. In addition, the use of this consortium enabled a rapid investigation of multiple pathways by inoculating different combinations of bacterial cells (2). To prepare the cell-free expression system, the researchers simply obtained cell lysates from the consortia without the need to purify and supplement individual proteins. The researchers first tested the protein expression capability of their consortia by measuring deGFP levels. They tested 18and 7-strain consortia expressing various initiation, elongation and termination factors, as well as aminoacyl-tRNA transferases, and found their expression levels to be comparable. In comparison to the wildtype bacteria and commercially available expression system (S30 T7 system), the consortium demonstrated &gt;2-fold increase in deGFP production. Interestingly, when the team investigated the underlying mechanism of the improvement, they found that simple addition of translation machineries did not fully explain the increase in protein synthesis. Thus, they hypothesized that the overexpression of translation machineries in cells led to host reprogramming of the proteome that favors protein synthesis. To investigate the shift in the proteome, the researchers analyzed protein composition using mass spectrometry. They found that the consortium indeed exhibited a global proteome shift compared to the controls, resulting in changes in the expression level of more than 700 proteins. Importantly, these changes were associated with upregulation ","PeriodicalId":74902,"journal":{"name":"Synthetic biology (Oxford, England)","volume":"5 1","pages":"ysaa013"},"PeriodicalIF":0.0,"publicationDate":"2020-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/synbio/ysaa013","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38436595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}