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BACKGROUND: Phenylpropanoids are a large group of plant secondary metabolites with various biological functions, derived from aromatic amino acids. Cyanobacteria are promising host organisms for sustainable production of plant phenylpropanoids. We have previously engineered Synechocystis sp. PCC 6803 to produce trans-cinnamic acid (tCA) and p-coumaric acid (pCou), the first intermediates of phenylpropanoid pathway, by overexpression of phenylalanine- and tyrosine ammonia lyases. In this study, we aimed to enhance the production of the target compounds tCA and pCou in Synechocystis. RESULTS: We eliminated the 4-hydroxyphenylpyruvate dioxygenase (HPPD) activity, which is a competing pathway consuming tyrosine and, possibly, phenylalanine for tocopherol synthesis. Moreover, several genes of the terminal steps of the shikimate pathway were overexpressed alone or in operons, such as aromatic transaminases, feedback insensitive cyclohexadienyl dehydrogenase (TyrC) from Zymomonas mobilis and the chorismate mutase (CM) domain of the fused chorismate mutase/prephenate dehydratase enzyme from Escherichia coli. The obtained engineered strains demonstrated nearly 1.5 times enhanced tCA and pCou production when HPPD was knocked out compared to the parental production strains, accumulating 138 ± 3.5 mg L-1 of tCA and 72.3 ± 10.3 mg L-1 of pCou after seven days of photoautotrophic growth. However, there was no further improvement when any of the pathway genes were overexpressed. Finally, we used previously obtained AtPRM8 and TsPRM8 Synechocystis strains with deregulated shikimate pathway as a background for the overexpression of synthetic constructs with ppd knockout. CONCLUSIONS: HPPD elimination enhances the tCA and pCou productivity to a similar extent. The use of PRM8 based strains as a background for overexpression of synthetic constructs, however, did not promote tCA and pCou titers, which indicates a tight regulation of the terminal steps of phenylalanine and tyrosine synthesis. This work contributes to establishing cyanobacteria as hosts for phenylpropanoid production.
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Synechocystis , Synechocystis/genética , Synechocystis/metabolismo , Ingeniería Metabólica , Ácido Shikímico/metabolismo , Tirosina/metabolismo , Fenilalanina/metabolismo , Corismato Mutasa/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismoRESUMEN
The looming climate crisis has prompted an ever-growing interest in cyanobacteria due to their potential as sustainable production platforms for the synthesis of energy carriers and value-added chemicals from CO2 and sunlight. Nonetheless, cyanobacteria are yet to compete with heterotrophic systems in terms of space-time yields and consequently production costs. One major drawback leading to the low production performance observed in cyanobacteria is the limited ability to utilize the full capacity of the photosynthetic apparatus and its associated systems, i.e. CO2 fixation and the directly connected metabolism. In this review, novel insights into various levels of metabolic regulation of cyanobacteria are discussed, including the potential of targeting these regulatory mechanisms to create a chassis with a phenotype favorable for photoautotrophic production. Compared to conventional metabolic engineering approaches, minor perturbations of regulatory mechanisms can have wide-ranging effects.
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Cianobacterias , Ingeniería Metabólica , Fotosíntesis , Ingeniería Metabólica/métodos , Cianobacterias/metabolismo , Cianobacterias/genética , Fotosíntesis/genética , Dióxido de Carbono/metabolismoRESUMEN
INTRODUCTION: Portfolio with a collection of evidence has become popular in higher education, including dental education. It is valuable to study the experiences of the use and implementation processes. Meta-ethnography can be a suitable method to analyse, synthesize and construct interpretations of qualitative research. Our aim was to explore experiences from the use of a portfolio/e-portfolio in dental education, from the students' and teachers' perspectives. MATERIALS AND METHODS: A systematic search in the databases PubMed, Scopus and ERC was performed, and the established seven steps of a meta-ethnographic review were used. 278 papers were initially identified, and seven were included in the final analysis. RESULTS: Two themes (Issues to Address and Consequences) and five subthemes (Purpose, Roles, Support and Structure, Challenges and Enablers, and Gains) were constructed. DISCUSSION: Our synthesis reflects various challenges, yet the learning gains are recognized and expressed to be important once the students and teachers have overcome early thresholds. Beyond the conclusions drawn in each paper, our synthesis provides new perspectives on the complexity of an implementation process and the balance of not seeing the woods for the trees being overwhelmed by technical and other practical aspects, reducing the opportunity for learning. CONCLUSION: The portfolio implementation in undergraduate dental education should address clarification to all stakeholders of the purpose and role, presenting a purposeful portfolio structure and timely support.
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Educación en Odontología , Investigación Cualitativa , Educación en Odontología/métodos , Humanos , Antropología Cultural , Estudiantes de Odontología/psicología , Evaluación Educacional/métodos , AprendizajeRESUMEN
Background: During the COVID-19 pandemic a need to process large volumes of publications emerged. As the pandemic is winding down, the clinicians encountered a novel syndrome - Post-acute Sequelae of COVID-19 (PASC) - that affects over 10 % of those who contract SARS-CoV-2 and presents a significant challenge in the medical field. The continuous influx of publications underscores a need for efficient tools for navigating the literature. Objectives: We aimed to develop an application which will allow monitoring and categorizing COVID-19-related literature through building publication networks and medical subject headings (MeSH) maps to identify key publications and networks. Methods: We introduce CORACLE (COVID-19 liteRAture CompiLEr), an innovative web application designed to analyse COVID-19-related scientific articles and to identify research trends. CORACLE features three primary interfaces: The "Search" interface, which displays research trends and citation links; the "Citation Map" interface, allowing users to create tailored citation networks from PubMed Identifiers (PMIDs) to uncover common references among selected articles; and the "MeSH" interface, highlighting current MeSH trends and their associations. Results: CORACLE leverages PubMed data to categorize literature on COVID-19 and PASC, aiding in the identification of relevant research publication hubs. Using lung function in PASC patients as a search example, we demonstrate how to identify and visualize the interactions between the relevant publications. Conclusion: CORACLE is an effective tool for the extraction and analysis of literature. Its functionalities, including the MeSH trends and customizable citation mapping, facilitate the discovery of emerging trends in COVID-19 and PASC research.