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1.
Proc Natl Acad Sci U S A ; 119(30): e2119872119, 2022 07 26.
Artículo en Inglés | MEDLINE | ID: mdl-35858416

RESUMEN

At present, there is no simple, first principles-based, and general model for quantitatively describing the full range of observed biological temperature responses. Here we derive a general theory for temperature dependence in biology based on Eyring-Evans-Polanyi's theory for chemical reaction rates. Assuming only that the conformational entropy of molecules changes with temperature, we derive a theory for the temperature dependence of enzyme reaction rates which takes the form of an exponential function modified by a power law and that describes the characteristic asymmetric curved temperature response. Based on a few additional principles, our model can be used to predict the temperature response above the enzyme level, thus spanning quantum to classical scales. Our theory provides an analytical description for the shape of temperature response curves and demonstrates its generality by showing the convergence of all temperature dependence responses onto universal relationships-a universal data collapse-under appropriate normalization and by identifying a general optimal temperature, around 25 ∘C, characterizing all temperature response curves. The model provides a good fit to empirical data for a wide variety of biological rates, times, and steady-state quantities, from molecular to ecological scales and across multiple taxonomic groups (from viruses to mammals). This theory provides a simple framework to understand and predict the impact of temperature on biological quantities based on the first principles of thermodynamics, bridging quantum to classical scales.


Asunto(s)
Fenómenos Biológicos , Temperatura , Animales , Biología , Mamíferos , Termodinámica , Virus
2.
Appl Environ Microbiol ; 88(1): e0184221, 2022 01 11.
Artículo en Inglés | MEDLINE | ID: mdl-34705547

RESUMEN

Polyethylene terephthalate (PET) is one of the most widely used synthetic plastics in the packaging industry, and consequently has become one of the main components of plastic waste found in the environment. However, several microorganisms have been described to encode enzymes that catalyze the depolymerization of PET. While most known PET hydrolases are thermophilic and require reaction temperatures between 60°C and 70°C for an efficient hydrolysis of PET, a partial hydrolysis of amorphous PET at lower temperatures by the polyester hydrolase IsPETase from the mesophilic bacterium Ideonella sakaiensis has also been reported. We show that polyester hydrolases from the Antarctic bacteria Moraxella sp. strain TA144 (Mors1) and Oleispira antarctica RB-8 (OaCut) were able to hydrolyze the aliphatic polyester polycaprolactone as well as the aromatic polyester PET at a reaction temperature of 25°C. Mors1 caused a weight loss of amorphous PET films and thus constitutes a PET-degrading psychrophilic enzyme. Comparative modeling of Mors1 showed that the amino acid composition of its active site resembled both thermophilic and mesophilic PET hydrolases. Lastly, bioinformatic analysis of Antarctic metagenomic samples demonstrated that members of the Moraxellaceae family carry candidate genes coding for further potential psychrophilic PET hydrolases. IMPORTANCE A myriad of consumer products contains polyethylene terephthalate (PET), a plastic that has accumulated as waste in the environment due to its long-term stability and poor waste management. One promising solution is the enzymatic biodegradation of PET, with most known enzymes only catalyzing this process at high temperatures. Here, we bioinformatically identified and biochemically characterized an enzyme from an Antarctic organism that degrades PET at 25°C with similar efficiency to the few PET-degrading enzymes active at moderate temperatures. Reasoning that Antarctica harbors other PET-degrading enzymes, we analyzed available data from Antarctic metagenomic samples and successfully identified other potential enzymes. Our findings contribute to increasing the repertoire of known PET-degrading enzymes that are currently being considered as biocatalysts for the biological recycling of plastic waste.


Asunto(s)
Hidrolasas , Tereftalatos Polietilenos , Regiones Antárticas , Hidrolasas/genética , Hidrólisis , Poliésteres , Temperatura
3.
Molecules ; 27(23)2022 Dec 03.
Artículo en Inglés | MEDLINE | ID: mdl-36500606

RESUMEN

Key organisms in the environment, such as oxygenic photosynthetic primary producers (photosynthetic eukaryotes and cyanobacteria), are responsible for fixing most of the carbon globally. However, they are affected by environmental conditions, such as temperature, which in turn affect their distribution. Globally, the cyanobacterium Fischerella thermalis is one of the main primary producers in terrestrial hot springs with thermal gradients up to 60 °C, but the mechanisms by which F. thermalis maintains its photosynthetic activity at these high temperatures are not known. In this study, we used molecular approaches and bioinformatics, in addition to photophysiological analyses, to determine the genetic activity associated with the energy metabolism of F. thermalis both in situ and in high-temperature (40 °C to 65 °C) cultures. Our results show that photosynthesis of F. thermalis decays with temperature, while increased transcriptional activity of genes encoding photosystem II reaction center proteins, such as PsbA (D1), could help overcome thermal damage at up to 60 °C. We observed that F. thermalis tends to lose copies of the standard G4 D1 isoform while maintaining the recently described D1INT isoform, suggesting a preference for photoresistant isoforms in response to the thermal gradient. The transcriptional activity and metabolic characteristics of F. thermalis, as measured by metatranscriptomics, further suggest that carbon metabolism occurs in parallel with photosynthesis, thereby assisting in energy acquisition under high temperatures at which other photosynthetic organisms cannot survive. This study reveals that, to cope with the harsh conditions of hot springs, F. thermalis has several compensatory adaptations, and provides emerging evidence for mixotrophic metabolism as being potentially relevant to the thermotolerance of this species. Ultimately, this work increases our knowledge about thermal adaptation strategies of cyanobacteria.


Asunto(s)
Cianobacterias , Electrones , Cianobacterias/genética , Cianobacterias/metabolismo , Fotosíntesis , Carbono/metabolismo
4.
Extremophiles ; 23(6): 635-647, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31512055

RESUMEN

The true-branching cyanobacterium Fischerella thermalis (also known as Mastigocladus laminosus) is widely distributed in hot springs around the world. Morphologically, it has been described as early as 1837. However, its taxonomic placement remains controversial. F. thermalis belongs to the same genus as mesophilic Fischerella species but forms a monophyletic clade of thermophilic Fischerella strains and sequences from hot springs. Their recent divergence from freshwater or soil true-branching species and the ongoing process of specialization inside the thermal gradient make them an interesting evolutionary model to study. F. thermalis is one of the most complex prokaryotes. It forms a cellular network in which the main trichome and branches exchange metabolites and regulators via septal junctions. This species can adapt to a variety of environmental conditions, with its photosynthetic apparatus remaining active in a temperature range from 15 to 58 °C. Together with its nitrogen-fixing ability, this allows it to dominate in hot spring microbial mats and contribute significantly to the de novo carbon and nitrogen input. Here, we review the current knowledge on the taxonomy and distribution of F. thermalis, its morphological complexity, and its physiological adaptations to an extreme environment.


Asunto(s)
Aclimatación/fisiología , Evolución Biológica , Cianobacterias/fisiología , Manantiales de Aguas Termales/microbiología , Calor , Modelos Biológicos , Tricomas/fisiología
5.
Rev Esp Enferm Dig ; 111(11): 887-888, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31595762

RESUMEN

Endometriosis is the presence of extra-uterine endometrial tissue and affects 6-10% of fertile women. In 3-37% of the cases it affects the gastrointestinal tract being the ileal location extremely infrequent. Symptoms are nonspecific, debuting as intestinal obstruction in 7-23% of cases. The definitive diagnosis is anatomopathological and the treatment in these cases of acute abdomen is surgery. We attach the present case due to its rarity, it is a young asymptomatic woman who debuts with a picture of intestinal obstruction that is treated by intestinal resection via laparoscopy confirming the endometriosis etiology.


Asunto(s)
Endometriosis/cirugía , Enfermedades del Íleon/cirugía , Obstrucción Intestinal/cirugía , Laparoscopía , Adulto , Endometriosis/complicaciones , Femenino , Humanos , Enfermedades del Íleon/complicaciones , Obstrucción Intestinal/etiología
6.
Pediatr Phys Ther ; 31(3): 272-278, 2019 07.
Artículo en Inglés | MEDLINE | ID: mdl-31225831

RESUMEN

PURPOSE: To report on the preliminary aquatic physical therapy core sets for children and youth with neurological disorders using the International Classification of Functioning, Disability and Health-children and youth version. METHODS: A formal and structured consensus process was developed, integrating the findings of preparatory studies: a systematic literature review, expert survey with 69 participants, and a qualitative study with 43 participants. RESULTS: Five preliminary aquatic physical therapy core sets were described: a Comprehensive Core Set (64 categories); a Common Brief (13 categories); and 3 age-specific Core Sets: infant (below 6 years, 18 categories), school-age (from 6 to <14 years, 22 categories), and youth (from 14 to 18 years, 19 categories). CONCLUSIONS: Consensus among aquatic physical therapists' expert opinions identified the relevant intervention categories available when treating children and youth with neurological disorders. This list of intervention categories can be used in practice, research, education, and health administration.


Asunto(s)
Enfermedades del Sistema Nervioso/rehabilitación , Fisioterapeutas/psicología , Modalidades de Fisioterapia , Agua , Actividades Cotidianas , Adolescente , Niño , Preescolar , Consenso , Evaluación de la Discapacidad , Personas con Discapacidad , Femenino , Humanos , Masculino , Investigación Cualitativa , Encuestas y Cuestionarios
7.
Rev Esp Enferm Dig ; 110(3): 209, 2018 03.
Artículo en Inglés | MEDLINE | ID: mdl-29332403

RESUMEN

Colonoscopy is the gold standard for the study of colorectal pathology. Splenic injury is a rare but potentially fatal complication to consider. Therefore, we present two cases whose management was different and we show their clinical presentation, their diagnosis and their treatment in order to recognize this complication early to establish early treatment.


Asunto(s)
Esplenectomía , Rotura del Bazo , Colonoscopía , Humanos
8.
Rev Esp Enferm Dig ; 110(4): 263, 2018 04.
Artículo en Inglés | MEDLINE | ID: mdl-29620412

RESUMEN

I have read Dr. Luzón's article (1) and have shared his opinions for a long time. As I indicated in my letter 13 years ago (2), all of our patients need to receive adequate sedation in endoscopic explorations, including ERCP. It is also our obligation as the person responsible for the procedure, to ensure that this is so. At present and without renouncing any of my beliefs, I think it is necessary to ask ourselves several questions: .


Asunto(s)
Endoscopía , Humanos
9.
Rev Esp Enferm Dig ; 109(4): 305-306, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-28112961

RESUMEN

Jejunal pseudodiverticulosis is an uncommon entity. Pseudodiverticulum are usually asymptomatic and an incidental finding. They can have a less frequent acute onset with perforation, obstruction or bleeding and they could have high morbidity and mortality. We report three patients who debuted with an acute abdomen.


Asunto(s)
Divertículo/diagnóstico , Obstrucción Intestinal/diagnóstico , Enfermedades del Yeyuno/diagnóstico , Anciano , Anciano de 80 o más Años , Diagnóstico Diferencial , Divertículo/diagnóstico por imagen , Femenino , Humanos , Hallazgos Incidentales , Obstrucción Intestinal/diagnóstico por imagen , Perforación Intestinal , Enfermedades del Yeyuno/diagnóstico por imagen , Masculino , Tomografía Computarizada por Rayos X
10.
J Physiol Biochem ; 80(1): 1-9, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38019451

RESUMEN

Hypothyroidism is the most frequent endocrine pathology. Although clinical or overt hypothyroidism has been traditionally associated to low T3 / T4 and high thyrotropin (TSH) circulating levels, other forms exist such as subclinical hypothyroidism, characterized by normal blood T3 / T4 and high TSH. In its different forms is estimated to affect approximately 10% of the population, especially women, in a 5:1 ratio with respect to men. Among its consequences are alterations in cardiac electrical activity, especially in the repolarization phase, which is accompanied by an increased susceptibility to cardiac arrhythmias. Although these alterations have traditionally been attributed to thyroid hormone deficiency, recent studies, both clinical trials and experimental models, demonstrate a fundamental role of TSH in cardiac electrical remodeling. Thus, both metabolic thyroid hormones and TSH regulate cardiac ion channel expression in many and varied ways. This means that the different combinations of hormones that predominate in different types of hypothyroidism (overt, subclinic, primary, central) can generate different forms of cardiac electrical remodeling. These new findings are raising the relevant question of whether serum TSH reference ranges should be redefined.


Asunto(s)
Remodelación Atrial , Hipotiroidismo , Masculino , Femenino , Humanos , Triyodotironina/uso terapéutico , Tirotropina/uso terapéutico , Tiroxina/uso terapéutico
11.
ISME Commun ; 4(1): ycae059, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38770060

RESUMEN

Mobile genetic elements (MGEs), collectively referred to as the "mobilome", can have a significant impact on the fitness of microbial communities and therefore on ecological processes. Marine MGEs have mainly been associated with wide geographical and phylogenetic dispersal of adaptative traits. However, whether the structure of this mobilome exhibits deterministic patterns in the natural community is still an open question. The aim of this study was to characterize the structure of the conjugative mobilome in the ocean surface bacterioplankton by searching the publicly available marine metagenomes from the TARA Oceans survey, together with molecular markers, such as relaxases and type IV coupling proteins of the type IV secretion system (T4SS). The T4SS machinery was retrieved in more abundance than relaxases in the surface marine bacterioplankton. Moreover, among the identified MGEs, mobilizable elements were the most abundant, outnumbering self-conjugative sequences. Detection of a high number of incomplete T4SSs provides insight into possible strategies related to trans-acting activity between MGEs, and accessory functions of the T4SS (e.g. protein secretion), allowing the host to maintain a lower metabolic burden in the highly dynamic marine system. Additionally, the results demonstrate a wide geographical dispersion of MGEs throughout oceanic regions, while the Southern Ocean appears segregated from other regions. The marine mobilome also showed a high similarity of functions present in known plasmid databases. Moreover, cargo genes were mostly related to DNA processing, but scarcely associated with antibiotic resistance. Finally, within the MGEs, integrative and conjugative elements showed wider marine geographic dispersion than plasmids.

12.
Front Microbiol ; 15: 1360268, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38633703

RESUMEN

Recent studies have expanded the genomic contours of the Acidithiobacillia, highlighting important lacunae in our comprehension of the phylogenetic space occupied by certain lineages of the class. One such lineage is 'Igneacidithiobacillus', a novel genus-level taxon, represented by 'Igneacidithiobacillus copahuensis' VAN18-1T as its type species, along with two other uncultivated metagenome-assembled genomes (MAGs) originating from geothermally active sites across the Pacific Ring of Fire. In this study, we investigate the genetic and genomic diversity, and the distribution patterns of several uncharacterized Acidithiobacillia class strains and sequence clones, which are ascribed to the same 16S rRNA gene sequence clade. By digging deeper into this data and contributing to novel MAGs emerging from environmental studies in tectonically active locations, the description of this novel genus has been consolidated. Using state-of-the-art genomic taxonomy methods, we added to already recognized taxa, an additional four novel Candidate (Ca.) species, including 'Ca. Igneacidithiobacillus chanchocoensis' (mCHCt20-1TS), 'Igneacidithiobacillus siniensis' (S30A2T), 'Ca. Igneacidithiobacillus taupoensis' (TVZ-G3 TS), and 'Ca. Igneacidithiobacillus waiarikiensis' (TVZ-G4 TS). Analysis of published data on the isolation, enrichment, cultivation, and preliminary microbiological characterization of several of these unassigned or misassigned strains, along with the type species of the genus, plus the recoverable environmental data from metagenomic studies, allowed us to identify habitat preferences of these taxa. Commonalities and lineage-specific adaptations of the seven species of the genus were derived from pangenome analysis and comparative genomic metabolic reconstruction. The findings emerging from this study lay the groundwork for further research on the ecology, evolution, and biotechnological potential of the novel genus 'Igneacidithiobacillus'.

13.
Extremophiles ; 17(1): 123-36, 2013 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-23208511

RESUMEN

Seasonal shifts in bacterial diversity of microbial mats were analyzed in three hot springs (39-68 °C) of Patagonia, using culture-independent methods. Three major bacterial groups were detected in all springs: Phyla Cyanobacteria and Bacteroidetes, and Order Thermales. Proteobacteria, Acidobacteria and Green Non-Sulfur Bacteria were also detected in small amounts and only in some samples. Thermophilic filamentous heterocyst-containing Mastigocladus were dominant Cyanobacteria in Porcelana Hot Spring and Geyser, and Calothrix in Cahuelmó, followed by the filamentous non-heterocyst Leptolyngbya and Oscillatoria. Bacteroidetes were detected in a wide temperature range and their relative abundance increased with decreasing temperature in almost all samples. Two Meiothermus populations with different temperature optima were found. Overall, fingerprinting analysis with universal bacterial primers showed high similarities within each hot spring despite differences in temperature. On the other hand, Cahuelmó Hot Spring showed a lower resemblance among samples. Porcelana Hot Spring and Porcelana Geyser were rather similar to each other, possibly due to a common geological substrate given their geographic proximity. This was even more evident with specific cyanobacterial primers. The different geological substrate and the seawater influence in Cahuelmó might have caused the differences in the microbial community structure with the other two hot springs.


Asunto(s)
Bacterias/crecimiento & desarrollo , Biodiversidad , Manantiales de Aguas Termales/microbiología , Microbiología del Agua , Bacterias/genética
14.
Microbiol Resour Announc ; 12(6): e0005923, 2023 Jun 20.
Artículo en Inglés | MEDLINE | ID: mdl-37184380

RESUMEN

Microbes play an important role in coastal and estuarine waters. We present 93 metagenomes and 677 metagenome-assembled genomes (MAGs) from Comau Fjord, Patagonia (42°S), to further understand the microbial dynamics and their response to anthropogenic disturbances. These data represent a spatially (35-km transect) and temporally (2016 to 2019) explicit data set.

15.
Microbiol Resour Announc ; 12(4): e0008223, 2023 Apr 18.
Artículo en Inglés | MEDLINE | ID: mdl-36946737

RESUMEN

Viruses are key players in marine environments, affecting food webs and biogeochemical cycles. We present 48 viral metagenomes and 5,656 viral operational taxonomic units (vOTUs) from Comau Fjord, Patagonia (42°S), to understand viral-mediated processes in coastal and estuarine waters. These data represent a spatial (35-km transect, two depths) and seasonal (winter and fall) data set.

16.
Microorganisms ; 11(4)2023 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-37110327

RESUMEN

While progress has been made in surveying the oceans to understand microbial and viral communities, the coastal ocean and, specifically, estuarine waters, where the effects of anthropogenic activity are greatest, remain partially understudied. The coastal waters of Northern Patagonia are of interest since this region experiences high-density salmon farming as well as other disturbances such as maritime transport of humans and cargo. Here, we hypothesized that viral and microbial communities from the Comau Fjord would be distinct from those collected in global surveys yet would have the distinctive features of microbes from coastal and temperate regions. We further hypothesized that microbial communities will be functionally enriched in antibiotic resistance genes (ARGs) in general and in those related to salmon farming in particular. Here, the analysis of metagenomes and viromes obtained for three surface water sites showed that the structure of the microbial communities was distinct in comparison to global surveys such as the Tara Ocean, though their composition converges with that of cosmopolitan marine microbes belonging to Proteobacteria, Bacteroidetes, and Actinobacteria. Similarly, viral communities were also divergent in structure and composition but matched known viral members from North America and the southern oceans. Microbial communities were functionally enriched in ARGs dominated by beta-lactams and tetracyclines, bacitracin, and the group macrolide-lincosamide-streptogramin (MLS) but were not different from other communities from the South Atlantic, South Pacific, and Southern Oceans. Similarly, viral communities were characterized by exhibiting protein clusters similar to those described globally (Tara Oceans Virome); however, Comau Fjord viromes displayed up to 50% uniqueness in their protein content. Altogether, our results indicate that microbial and viral communities from the Comau Fjord are a reservoir of untapped diversity and that, given the increasing anthropogenic impacts in the region, they warrant further study, specifically regarding resilience and resistance against antimicrobials and hydrocarbons.

17.
Am J Infect Control ; 51(12): 1324-1328, 2023 12.
Artículo en Inglés | MEDLINE | ID: mdl-37295678

RESUMEN

BACKGROUND: Our aim was to evaluate the effectiveness of an intervention to reduce the incidence of non-ventilator-associated hospital-acquired pneumonia (NV-HAP) and determine compliance with preventive measures. METHODS: This was a quasi-experimental before-after study involving patients in the 53-bed Internal Medicine ward in a university hospital in Spain. The preventive measures included hand hygiene, dysphagia detection, head-of-bed elevation, withdrawal of sedatives in the event of confusion, oral care, and sterile or bottled water use. A prospective post-intervention study of the incidence of NV-HAP was carried out from February 2017 to January 2018 and compared with baseline incidence (May 2014 to April 2015). Compliance with preventive measures was analyzed with 3-point-prevalence studies (December 2015, October 2016, and June 2017). RESULTS: The rate of NV-HAP decreased from 0.45 cases (95% confidence interval 0.24-0.77) in the pre-intervention period to 0.18 cases per 1,000 patient-days (95% confidence interval 0.07-0.39) in the post-intervention period (P = .07). Compliance with most preventive measures improved after intervention and remained stable over time. CONCLUSIONS: The strategy improved the adherence to most of the preventive measures, with a decrease in the incidence of NV-HAP. Efforts to enhance adherence to such fundamental preventive measures are critical to lowering the incidence of NV-HAP.


Asunto(s)
Neumonía Asociada al Ventilador , Humanos , Proyectos Piloto , Incidencia , Estudios Prospectivos , Neumonía Asociada al Ventilador/epidemiología , Neumonía Asociada al Ventilador/prevención & control , Hospitales Universitarios
18.
Microbiol Spectr ; : e0024923, 2023 Sep 27.
Artículo en Inglés | MEDLINE | ID: mdl-37754764

RESUMEN

Temperature, pH, and hydrochemistry of terrestrial hot springs play a critical role in shaping thermal microbial communities. However, the interactions of biotic and abiotic factors at this terrestrial-aquatic interface are still not well understood on a global scale, and the question of how underground events influence microbial communities remains open. To answer this, 11 new samples obtained from the El Tatio geothermal field were analyzed by 16S rRNA amplicon sequencing (V4 region), along with 191 samples from previous publications obtained from the Taupo Volcanic Zone, the Yellowstone Plateau Volcanic Field, and the Eastern Tibetan Plateau, with their temperature, pH, and major ion concentration. Microbial alpha diversity was lower in acid-sulfate waters, and no significant correlations were found with temperature. However, moderate correlations were observed between chemical parameters such as pH (mostly constrained to temperatures below 70°C), SO4 2- and abundances of members of the phyla Armatimonadota, Deinococcota, Chloroflexota, Campilobacterota, and Thermoplasmatota. pH and SO4 2- gradients were explained by phase separation of sulfur-rich hydrothermal fluids and oxidation of reduced sulfur in the steam phase, which were identified as key processes shaping these communities. Ordination and permutational analysis of variance showed that temperature, pH, and major element hydrochemistry explain only 24% of the microbial community structure. Therefore, most of the variance remained unexplained, suggesting that other environmental or biotic factors are also involved and highlighting the environmental complexity of the ecosystem and its great potential to test niche theory ecological associated questions. IMPORTANCE This is the first approach to investigate whether geothermal processes could have an influence on the ecology of thermal microbial communities on a global scale. In addition to temperature and pH, microbial communities are structured by sulfate concentrations, which depends on the tectono-magmatic settings (such as the depth of magmatic chambers) and the local settings (such as the availability of a confining layer separating NaCl waters from steam after phase separation) and the possibility of mixing with more diluted fluids. Comparison of microbial communities from different geothermal areas by homogeneous sequence processing showed that no significant geographic distance decay was detected on the microbial communities according to Bray-Curtis, Jaccard, unweighted, and weighted Unifrac similarity/dissimilarity indices. Instead, an ancient potential divergence in the same taxonomic groups is suggested between globally distant thermal zones.

19.
Microorganisms ; 11(7)2023 Jul 14.
Artículo en Inglés | MEDLINE | ID: mdl-37512975

RESUMEN

In terrestrial hot springs, some members of the microbial mat community utilize sulfur chemical species for reduction and oxidization metabolism. In this study, the diversity and activity of sulfur-metabolizing bacteria were evaluated along a temperature gradient (48-69 °C) in non-acidic phototrophic mats of the Porcelana hot spring (Northern Patagonia, Chile) using complementary meta-omic methodologies and specific amplification of the aprA (APS reductase) and soxB (thiosulfohydrolase) genes. Overall, the key players in sulfur metabolism varied mostly in abundance along the temperature gradient, which is relevant for evaluating the possible implications of microorganisms associated with sulfur cycling under the current global climate change scenario. Our results strongly suggest that sulfate reduction occurs throughout the whole temperature gradient, being supported by different taxa depending on temperature. Assimilative sulfate reduction is the most relevant pathway in terms of taxonomic abundance and activity, whereas the sulfur-oxidizing system (Sox) is likely to be more diverse at low rather than at high temperatures. Members of the phylum Chloroflexota showed higher sulfur cycle-related transcriptional activity at 66 °C, with a potential contribution to sulfate reduction and oxidation to thiosulfate. In contrast, at the lowest temperature (48 °C), Burkholderiales and Acetobacterales (both Pseudomonadota, also known as Proteobacteria) showed a higher contribution to dissimilative sulfate reduction/oxidation as well as to thiosulfate metabolism. Cyanobacteriota and Planctomycetota were especially active in assimilatory sulfate reduction. Analysis of the aprA and soxB genes pointed to members of the order Burkholderiales (Gammaproteobacteria) as the most dominant and active along the temperature gradient for these genes. Changes in the diversity and activity of different sulfur-metabolizing bacteria in photoautotrophic microbial mats along a temperature gradient revealed their important role in hot spring environments, especially the main primary producers (Chloroflexota/Cyanobacteriota) and diazotrophs (Cyanobacteriota), showing that carbon, nitrogen, and sulfur cycles are highly linked in these extreme systems.

20.
Protein Sci ; 32(9): e4757, 2023 09.
Artículo en Inglés | MEDLINE | ID: mdl-37574805

RESUMEN

Several hydrolases have been described to degrade polyethylene terephthalate (PET) at moderate temperatures ranging from 25°C to 40°C. These mesophilic PET hydrolases (PETases) are less efficient in degrading this plastic polymer than their thermophilic homologs and have, therefore, been the subject of many protein engineering campaigns. However, enhancing their enzymatic activity through rational design or directed evolution poses a formidable challenge due to the need for exploring a large number of mutations. Additionally, evaluating the improvements in both activity and stability requires screening numerous variants, either individually or using high-throughput screening methods. Here, we utilize instead the design of chimeras as a protein engineering strategy to increase the activity and stability of Mors1, an Antarctic PETase active at 25°C. First, we obtained the crystal structure of Mors1 at 1.6 Å resolution, which we used as a scaffold for structure- and sequence-based chimeric design. Then, we designed a Mors1 chimera via loop exchange of a highly divergent active site loop from the thermophilic leaf-branch compost cutinase (LCC) into the equivalent region in Mors1. After restitution of an active site disulfide bond into this chimera, the enzyme exhibited a shift in optimal temperature for activity to 45°C and an increase in fivefold in PET hydrolysis when compared with wild-type Mors1 at 25°C. Our results serve as a proof of concept of the utility of chimeric design to further improve the activity and stability of PETases active at moderate temperatures.


Asunto(s)
Hidrolasas , Tereftalatos Polietilenos , Tereftalatos Polietilenos/química , Tereftalatos Polietilenos/metabolismo , Regiones Antárticas , Hidrolasas/química , Hidrólisis , Ingeniería de Proteínas , Plásticos
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