Publications by authors named "Hugo Gramajo"

Article Synopsis
  • Over two decades, initiatives have aimed to enhance STEM undergraduate outcomes, with the inclusive Research Education Community (iREC) emerging as a scalable reform model that supports STEM faculty in implementing course-based research to improve student learning.
  • This study utilized pathway modeling to describe the HHMI Science Education Alliance (SEA) iREC, identifying how faculty engagement leads to sustainable adoption and improvement of new teaching strategies through feedback from over 100 participating faculty members.
  • The findings indicate that iREC fosters a collaborative environment where STEM faculty can share expertise and data, thereby enhancing their teaching practices and contributing to the overall evolution of undergraduate science education.
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PII proteins are signal transduction proteins that belong to a widely distributed family of proteins involved in the modulation of different metabolisms in bacteria. These proteins are homotrimers carrying a flexible loop, named T-loop, which changes its conformation due to the recognition of diverse key metabolites, ADP, ATP, and 2-oxoglutarate. PII proteins interact with different partners to primarily regulate a set of nitrogen pathways.

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Sustainable agriculture based on the use of soil-beneficial microbes such as plant growth-promoting rhizobacteria (PGPR) and biocontrol agents (BCA) is gaining great consideration to reduce the use of agrochemicals for crop production. With this aim, in this study, a total of 78 actinobacteria were isolated from the rhizosphere and endosphere of soybean roots. Based on in vitro compatibility with Bradyrhizobium japonicum, the ability to produce phytohormones, siderophores, exo-enzymes, antifungal compounds and phosphate solubilization (PGPR traits), two endophytic strains, named N2A and N9, were selected to evaluate their effects on plant growth and development at greenhouse and field conditions.

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In this work, we report the discovery and characterization of Garey24, a bacteriophage that forms medium-size plaques with halo rings isolated from a soil sample in Funes, Argentina. Its 41,522 bp circularly permuted genome contains 63 putative protein-coding genes. Based on gene content similarity, Garey24 was assigned to subcluster EA1.

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Article Synopsis
  • The study investigates the ability of marine microorganisms in coastal sediments to produce wax esters (WE) and triacylglycerols (TAG), which are important types of neutral lipids.
  • Researchers collected and analyzed sediment samples from subantarctic and Antarctic environments, finding a high abundance and diversity of bacteria capable of synthesizing these lipids, specifically looking at the gene associated with the key enzyme WS/DGAT.
  • The findings suggest that the observed lipid synthesis is linked to broader metabolic processes like fatty-acid metabolism and carbon recycling, highlighting the ecological significance of these bacteria in marine ecosystems.
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  • R. jostii RHA1 contains a diverse range of acyl-CoA carboxylases, including putative essential and non-essential ACC subunits involved in lipid biosynthesis.
  • Mutant strains lacking specific ACC genes showed that while some enzymes are non-essential for growth, RO04222 plays a significant role in the accumulation of lipids such as triacylglycerol.
  • The findings demonstrate the functional diversity and the varying essentiality of ACCs in actinobacteria, influenced by environmental conditions like nitrogen availability.
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Mycobacterial cell elongation occurs at the cell poles; however, it is not clear how cell wall insertion is restricted to the pole or how it is organized. Wag31 is a pole-localized cytoplasmic protein that is essential for polar growth, but its molecular function has not been described. In this study we used alanine scanning mutagenesis to identify Wag31 residues involved in cell morphogenesis.

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Background: The overprescription and misuse of classical antimicrobial compounds to treat gastrointestinal or systemic salmonellosis have been accelerating the surge of antibiotic-recalcitrant bacterial populations, posing a major public health challenge. Therefore, alternative therapeutic approaches to treat Salmonella infections are urgently required.

Objectives: To identify and characterize actinobacterial secreted compounds with inhibitory properties against the Salmonella enterica PhoP/PhoQ signal transduction system, crucial for virulence regulation.

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Background: A broad diversity of natural and non-natural esters have now been made in bacteria, and in other microorganisms, as a result of original metabolic engineering approaches. However, the fact that the properties of these molecules, and therefore their applications, are largely defined by the structural features of the fatty acid and alcohol moieties, has driven a persistent interest in generating novel structures of these chemicals.

Results: In this research, we engineered Escherichia coli to synthesize de novo esters composed of multi-methyl-branched-chain fatty acids and short branched-chain alcohols (BCA), from glucose and propionate.

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Article Synopsis
  • Phosphatidic acid phosphatase (PAP) converts phosphatidic acid into diacylglycerol, influencing the synthesis of triacylglycerol and glycerophospholipids.
  • Disrupting the two PAP genes in Mycobacterium smegmatis reduces TAG production during early growth but restores it when the bacteria enter exponential phase, despite lower phospholipid synthesis.
  • The changes in lipid composition in the PAP mutant, marked by increased odd chain fatty acids and impaired biofilm formation, highlight the importance of PAP in regulating lipid metabolism and maintaining lipid homeostasis, which is crucial for understanding Mycobacterium tuberculosis infection.
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  • Surfactin is a lipoheptapeptide discovered in 1969, synthesized by nonribosomal peptide synthetases, allowing for diversity among its peptide structures.
  • It can have fifteen different fatty acid isoforms (C12 to C17), leading to a variety of surfactin-like biomolecules with unique properties and biological activities.
  • Recent advancements in metabolic modeling, synthetic biology, and production optimization are aimed at enhancing surfactin biosynthesis and increasing cell productivity throughout its entire production process.
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In Mycobacterium tuberculosis, heparin-binding hemagglutinin (HBHAMT) has a relevant role in infection. It is also present in non-virulent mycobacteria and ancient actinobacteria, such as Rhodococcus opacus. To have a better understanding of the underlying mechanisms that shaped the evolutionary divergence of these proteins, we performed a comprehensive phylogenetic analysis of the regulatory sequences that drive the expression of hbha in saprophytic and pathogenic mycobacterial species.

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, the etiologic agent of human tuberculosis, is the world's leading cause of death from an infectious disease. One of the main features of this pathogen is the complex and dynamic lipid composition of the cell envelope, which adapts to the variable host environment and defines the fate of infection by actively interacting with and modulating immune responses. However, while much has been learned about the enzymes of the numerous lipid pathways, little knowledge is available regarding the proteins and metabolic signals regulating lipid metabolism during infection.

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Plant waxes are interesting substitutes of fossil-derived compounds; however, their limited sources and narrow structural diversity prompted the development of microbial platforms to produce esters with novel chemical structures and properties. One successful strategy was the heterologous expression of the mycocerosic polyketide synthase-based biosynthetic pathway (MAS-PKS, PapA5 and FadD28 enzymes) from Mycobacterium tuberculosis in Escherichia coli. This recombinant strain has the ability to produce a broad spectrum of multimethyl-branched long-chain esters (MBE) with novel chemical structures and high oxidation stability.

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Article Synopsis
  • Mycobacterium tuberculosis has a unique cell envelope structure with long fatty acids that contribute to its resistance and ability to modulate the host's immune response.
  • FasR is a key transcriptional activator that senses these fatty acids and regulates lipid biosynthesis in the bacteria.
  • Researchers revealed the crystal structures of FasR interacting with acyl ligands and DNA, showing how its structure changes to sense fatty acids and control genetic activity through a long tunnel that allows for effector binding and alters its configuration for DNA dissociation.
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Acetyl-CoA carboxylases (ACCs) are enzyme complexes generally composed of three catalytic domains and distributed in all organisms. In prokaryotes and plastids of most plants, these domains are encoded in distinct subunits forming heteromeric complexes. Distinctively, cytosolic ACCs from eukaryotes and plastids of graminaceous monocots, are organized in a single multidomain polypeptide.

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Iterative type I polyketide synthases (PKS) are megaenzymes essential to the biosynthesis of an enormously diverse array of bioactive natural products. Each PKS contains minimally three functional domains, β-ketosynthase (KS), acyltransferase (AT), and acyl carrier protein (ACP), and a subset of reducing domains such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER). The substrate selection, condensation reactions, and β-keto processing of the polyketide growing chain are highly controlled in a programmed manner.

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The development of new heterologous hosts for polyketides production represents an excellent opportunity to expand the genomic, physiological, and biochemical backgrounds that better fit the sustainable production of these valuable molecules. Cyanobacteria are particularly attractive for the production of natural compounds because they have minimal nutritional demands and several strains have well established genetic tools. Using the model strain Synechococcus elongatus, a generic platform was developed for the heterologous production of polyketide synthase (PKS)-derived compounds.

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Citrus canker is a disease caused by the phytopathogen Xanthomonas citri subsp. citri (Xcc), bacterium which is unable to survive out of the host for extended periods of time. Once established inside the plant, the pathogen must compete for resources and evade the defenses of the host cell.

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The complex lipids present in the cell wall of Mycobacterium tuberculosis (Mtb) act as major effector molecules that actively interact with the host, modulating its metabolism and stimulating the immune response, which in turn affects the physiology of both, the host cell and the bacilli. Lipids from the host are also nutrient sources for the pathogen and define the fate of the infection by modulating lipid homeostasis. Although new technologies and experimental models of infection have greatly helped understanding the different aspects of the host-pathogen interactions at the lipid level, the impact of this interaction in the Mtb lipid regulation is still incipient, mainly because of the low background knowledge in this area of research.

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Article Synopsis
  • The PII protein family is a highly conserved group of signal transduction proteins important for nitrogen and carbon metabolism, mainly studied in Gram-negative bacteria.
  • There is limited understanding of PII proteins from Gram-positive bacteria, particularly from Actinobacteria significant in medicine and biotechnology.
  • This study details successful methods for expressing and purifying untagged PII proteins from Actinobacteria, confirming their correct folding and paving the way for further biochemical research on their functions.
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Oleaginous microorganisms represent possible platforms for the sustainable production of oleochemicals and biofuels due to their metabolic robustness and the possibility to be engineered. is among the narrow group of prokaryotes capable of accumulating triacylglycerol (TAG) as carbon and energy reserve. Although the pathways for TAG biosynthesis in this organism have been widely addressed, the set of genes required for their breakdown have remained elusive so far.

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One of the dominant features of the biology of , and other mycobacteria, is the mycobacterial cell envelope with its exceptional complex composition. Mycolic acids are major and very specific components of the cell envelope and play a key role in its architecture and impermeability. Biosynthesis of mycolic acid (MA) precursors requires two types of fatty acid synthases, FAS I and FAS II, which should work in concert in order to keep lipid homeostasis tightly regulated.

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Conventional petroleum-based chemical industry, although economically still thriving, is now facing great socio-political challenges due to the increasing concerns on climate change and limited availability of fossil resources. In this context, microbial production of fuels and commodity oleochemicals from renewable biomass is being considered a promising sustainable alternative. The increasing understanding of cellular systems has enabled the redesign of microbial metabolism for the production of compounds present in many daily consumer products such as esters, waxes, fatty acids (FA) and fatty alcohols.

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