Diego Puente, Teresa de

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Diego Puente, Teresa de

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Diego Puente, Teresa de

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Universidad de Murcia. Departamento de Bioquímica y Biología Molecular"B" e Inmunología

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Open Access
Bacterial sirtuins overview: an open niche to explore
(Frontiers Media, 2021-10-26) Lozano Terol, Gema; Cánovas Díaz, Manuel; Diego Puente, Teresa de; Gallego Jara, Julia; Ortega Retuerta, Álvaro; Sola Martínez, Rosa Alba; Bioquímica y Biología Molecular B e Inmunología
Sirtuins are deacetylase enzymes widely distributed in all domains of life. Although for decades they have been related only to histones deacetylation in eukaryotic organisms, today they are considered global regulators in both prokaryotes and eukaryotes. Despite the important role of sirtuins in humans, the knowledge about bacterial sirtuins is still limited. Several proteomics studies have shown that bacterial sirtuins deacetylate a large number of lysines in vivo, although the effect that this deacetylation causes in most of them remains unknown. To date, only the regulation of a few bacterial sirtuin substrates has been characterized, being their metabolic roles widely distributed: carbon and nitrogen metabolism, DNA transcription, protein translation, or virulence. One of the most current topics on acetylation and deacetylation focuses on studying stoichiometry using quantitative LC-MS/MS. The results suggest that prokaryotic sirtuins deacetylate at low stoichiometry sites, although more studies are needed to know if it is a common characteristic of bacterial sirtuins and its biological significance. Unlike eukaryotic organisms, bacteria usually have one or few sirtuins, which have been reported to have closer common ancestors with the human Sirt5 than with any other class. In this work, in addition to carrying out an in-depth review of the role of bacterial sirtuins in their physiology, a phylogenetic study has been performed that reveals the evolutionary differences between sirtuins of different bacterial species and even between homologous sirtuins.
Open Access
Dynamic Lysine Acetylation Disrupts Isocitrate Lyase Function and Enables Metabolic Optimisation
(Wiley, 2026-03-30) Martínez Vivancos, Adrián; Gomariz-Turpin, Beatriz; Ortega Retuerta, Álvaro; Lozano Terol, Gema; Sola Martínez, Rosa Alba; Gallego Jara, Julia; Diego Puente, Teresa de; Bioquímica y Biología Molecular B e Inmunología
Proteomic studies have suggested that Escherichia coli isocitrate lyase (ICL) undergoes multiple acetylation events, partially inhibiting its activity. However, the molecular basis of this regulation and the contribution of individual lysine residues had not been defined. This study demonstrates that acetylation of ICL in E. coli is acetyl-phosphate–dependent and reversible by the CobB deacetylase, establishing a key post-translational regulatory mechanism within the glyoxylate shunt. Site-specific acetylation at K13 and K308 inhibits ICL activity by destabilising the tetrameric assembly and rendering the protein more prone to degradation, whereas lysine-to-arginine substitutions at these positions alleviate this inhibition, enhancing carbon flux distribution, metabolic flexibility and biomass yield without the burden of plasmid-based overexpression. Leveraging this regulatory insight, a KR mutant bearing lysine-to-arginine substitutions at residues 13 and 308, engineered directly into the chromosomal aceA gene, maintained wild-type growth rates while reducing acetate overflow and improving metabolic balance during glucose depletion and acetate assimilation, leading to a 61% increase in lycopene production. These findings highlight regulatory-based metabolic engineering as a powerful strategy to optimise bioproduction and pave the way for extending this approach to other central metabolic enzymes to develop robust microbial cell factories for the sustainable synthesis of biofuels, biochemicals and high-value compounds.
Open Access
The protein acetyltransferase PatZ from Escherichia coli is regulated by autoacetylation-induced oligomerization
(2015) Castaño Cerezo, Sara; Bernal Sanchez, Vicente; Manjón Rubio, Antonio; Fernández Espín, Vanesa; García de la Torre, José; Cánovas Díaz, Manuel; Diego Puente, Teresa de; Gallego Jara, Julia; Bioquímica y Biología Molecular B e Inmunología
Background: PatZ is the main Escherichia coli acetyltransferase and control acetyl-CoA synthetase (Acs) activity. Results: The kinetic and structural PatZ oligomer characteristics were determined. Conclusion: PatZ is a stable tetramer and forms an active octamer by autoacetylation to increase its stability. Significance: PTMs by acetylation have structural and functional roles in the cell.
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Engineering of microbial cell factories for production of plant-based natural products
(Elsevier, 2021) Gallego Jara, Julia; Lozano Terol, Gema; Sola Martínez, Rosa Alba; Cánovas Díaz, Manuel; Diego Puente, Teresa de; Bioquímica y Biología Molecular B e Inmunología; Facultades de la UMU::Facultad de Medicina
Biotechnology has become a promising alternative to produce highly valuable products. Production using microorganisms competes with chemical synthesis and extraction from natural sources, leading to cheaper and more sustainable production. To produce natural plant products using microorganisms, the development of molecular biology techniques that allow us to genetically and metabolically modify host microorganisms is essential. Thus many biotechnological processes have been developed to obtain terpenes, alkaloids, or polyphenols with interesting applications in the pharmaceutical, food, or cosmetic industries. The increase in our knowledge regarding the metabolism of host organisms, together with the continuous development of genetic and metabolic engineering techniques, will allow, in the coming years, biotechnology to be positioned as the main way to obtain high-value plant natural products.
Open Access
Data preprocessing workflow for exhaled breath analysis by GC/MS using open sources
(2020) Pastor Hernández, José María; Lozano-Terol, Gema; Cánovas Díaz, Manuel; Diego Puente, Teresa de; Gallego Jara, Julia; Sola Martínez, Rosa Alba; García-Marcos Álvarez, Luis Vicente; Bioquímica y Biología Molecular B e Inmunología
The noninvasive diagnosis and monitoring of high prevalence diseases such as cardiovascular diseases, cancers and chronic respiratory diseases are currently priority objectives in the area of health. In this regard, the analysis of volatile organic compounds (VOCs) has been identified as a potential noninvasive tool for the diagnosis and surveillance of several diseases. Despite the advantages of this strategy, it is not yet a routine clinical tool. The lack of reproducible protocols for each step of the biomarker discovery phase is an obstacle of the current state. Specifically, this issue is present at the data preprocessing step. Thus, an open source workflow for preprocessing the data obtained by the analysis of exhaled breath samples using gas chromatography coupled with single quadrupole mass spectrometry (GC/MS) is presented in this paper. This workflow is based on the connection of two approaches to transform raw data into a useful matrix for statistical analysis. Moreover, this workflow includes matching compounds from breath samples with a spectral library. Three free packages (xcms, cliqueMS and eRah) written in the language R are used for this purpose. Furthermore, this paper presents a suitable protocol for exhaled breath sample collection from infants under 2 years of age for GC/MS.
Open Access
Bacterial sirtuins overview: an open niche to explore
(Frontiers, 2021-10-26) Lozano Terol, Gema; Cánovas Díaz, Manuel; Diego Puente, Teresa de; Gallego Jara, Julia; Ortega Retuerta, Álvaro; Sola Martínez, Rosa Alba; Bioquímica y Biología Molecular B e Inmunología
Open Access
Comunicaciones Orales.- Structural analysis with DSC and Fluorescence in different mutants in lysine 101 of Catabolite Regulator Protein from Escherichia coli.
(2020-05-05) Écija Conesa, Ana; Arturo, Manjón Rubio; Cánovas Díaz, Manuel; Diego Puente, Teresa de; Gallego Jara, Julia; Facultades, Departamentos, Servicios y Escuelas::Departamentos de la UMU::Bioquímica y Biología Molecular B e Inmunología
Open Access
Impact of environmental exposures on exhaled breath and lung function: NELA Birth Cohort
(ERS publications, 2025) Jiménez Guerrero, Pedro; Sánchez-Solís de Querol, Manuel; Lozano Terol, Gema; Gallego Jara, Julia; Martínez Vivancos, Adrián; Morales Bartolomé, Eva; García-Marcos Álvarez, Luis Vicente; Diego Puente, Teresa de; NELA Study Group; Sola Martínez, Rosa Alba; Cirugía, Pediatría y Obstetricia y Ginecología; Facultad de Medicina
Introduction: Exposure to environmental factors (i.e. air pollution and second-hand tobacco smoke) have been associated with impaired lung function. However, the impact of environmental factors on lung health is usually evaluated separately and not with an exposomic framework. In this regard, breath analysis could be a noninvasive tool for biomonitoring of global human environmental exposure. Methods: Data come from 337 mother-child pairs from the Nutrition in Early Childhood Asthma (NELA) birth cohort. Levels of BTEX (benzene, toluene, ethylbenzene and xylenes) in exhaled breath from mothers and children at 3 months after birth were estimated using gas hromatography-mass spectrometry. Short-term residential exposures (breath sampling day and 15 days before breath sampling) to nitrogen dioxide, particulate matter (PM2.5) and ozone were determined by chemical dispersion/transport modelling. Forced vital capacity, forced expiratory volume in 0.5 s (FEV0.5) and forced expiratory flow at 75% of FVC and at 25%-75% of FVC were measured in infants according to the raised-volume rapid thoracoabdominal compression technique. Results: The results showed significant associations between short-term exposure to external agents and levels of benzene and toluene in exhaled breath. It was observed that exhaled levels of benzene and toluene were influenced by smoking status and outdoor air pollution in mothers, and by air pollution in infants (3 months of age). No significant relationship was observed between exposure to maternal tobacco smoking and/or short-term air pollution and lung function in healthy infants. However, there was a significant relationship between FEV0.5 and exhaled toluene in children. Discussion: These findings indicated a significant relationship between environmental exposures and exhaled levels of benzene and toluene, suggesting that breath analysis could be a helpful exposure biomonitoring tool.
Open Access
An ideal spacing is required for the control of Class II CRP-dependent promoters by the status of CRP K100
(2020-11-11) Ecija Conesa, Ana; Lozano-Terol, Gema; Browning, Douglas F.; Busby, Steve J.W.; Wolfe, Alan J.; Cánovas Díaz, Manuel; Diego Puente, Teresa de; Gallego Jara, Julia; Bioquímica y Biología Molecular B e Inmunología
Transcription activation by the Escherichia coli CRP at Class II promoters is dependent on direct interactions between RNA polymerase and CRP, therefore the spatial proximity between both proteins plays a significant role in the ability of CRP to activate transcription. Using both in vivo and in vitro techniques, here we demonstrate that the CRP K100 positive charge, adjacent to AR2, is required for full promoter activity when CRP is optimally positioned. Accordingly, K100 mediated activation is very position-dependent and our data confirm that the largest impact of the K100 status on transcription activation occurs when the spacing between the CRP binding site and the A2 of the -10 element is 22 bp. From the results of this study and the progress in the understanding about open complex DNA scrunching, we propose that CRP-dependent promoters should now be numbered by the distance from the centre of the DNA site for CRP and the most highly conserved base at position 2 of the -10 hexamer in bacterial promoters.
Open Access
Engineering protein production by rationally choosing a carbon and nitrogen source using E. coli BL21 acetate metabolism knockout strains.
(Springer Nature, 2019-09-04) Lozano Terol, Gema; Cánovas Díaz, Manuel; Diego Puente, Teresa de; Gallego Jara, Julia; Sola Martínez, Rosa Alba; Bioquímica y Biología Molecular B e Inmunología
Background: Escherichia coli (E. coli) is a bacteria that is widely employed in many industries for the production of high interest bio-products such as recombinant proteins. Nevertheless, the use of E. coli for recombinant protein production may entail some disadvantages such as acetate overfow. Acetate is accumulated under some culture conditions, involves a decrease in biomass and recombinant protein production, and its metabolism is related to protein lysine acetylation. Thereby, the carbon and nitrogen sources employed are relevant factors in cell host metabolism, and the study of the central metabolism of E. coli and its regulation is essential for optimizing the production of biomass and recombinant proteins. In this study, our aim was to fnd the most favourable conditions for carrying out recombinant protein production in E. coli BL21 using two diferent approaches, namely, manipulation of the culture media composition and the deletion of genes involved in acetate metabolism and Nε-lysine acetylation. Results: We evaluated protein overexpression in E. coli BL21 wt and fve mutant strains involved in acetate metabolism (Δacs, ΔackA and Δpta) and lysine acetylation (ΔpatZ and ΔcobB) grown in minimal medium M9 (inorganic ammonium nitrogen source) and in complex TB7 medium (peptide-based nitrogen source) supplemented with glucose (PTS carbon source) or glycerol (non-PTS carbon source). We observed a dependence of recombinant protein production on acetate metabolism and the carbon and nitrogen source employed. The use of complex medium supplemented with glycerol as a carbon source entails an increase in protein production and an efcient use of resources, since is a sub-product of biodiesel synthesis. Furthermore, the deletion of the ackA gene results in a fvefold increase in protein production with respect to the wt strain and a reduction in acetate accumulation. Conclusion: The results showed that the use of diverse carbon and nitrogen sources and acetate metabolism knockout strains can redirect E. coli carbon fuxes to diferent pathways and afect the fnal yield of the recombinant protein bioprocess. Thereby, we obtained a fvefold increase in protein production and an efcient use of the resources employing the most suitable strain and culture conditions.