A novel programmable lysozyme-based lysis system in Pseudomonas putida for biopolymer production

Indexación: Scopus; Web of Science.Cell lysis is crucial for the microbial production of industrial fatty acids, proteins, biofuels, and biopolymers. In this work, we developed a novel programmable lysis system based on the heterologous expression of lysozyme. The inducible lytic system was tested in two Gram-negative bacterial strains, namely Escherichia coli and Pseudomonas putida KT2440. Before induction, the lytic system did not significantly arrest essential physiological parameters in the recombinant E. coli (ECPi) and P. putida (JBOi) strain such as specific growth rate and biomass yield under standard growth conditions. A different scenario was observed in the recombinant JBOi strain when subjected to PHA-producing conditions, where biomass production was reduced by 25% but the mcl-PHA content was maintained at about 30% of the cell dry weight. Importantly, the genetic construct worked well under PHA-producing conditions (nitrogen-limiting phase), where more than 95% of the cell population presented membrane disruption 16 h post induction, with 75% of the total synthesized biopolymer recovered at the end of the fermentation period. In conclusion, this new lysis system circumvents traditional, costly mechanical and enzymatic cell-disrupting procedures.https://www.nature.com/articles/s41598-017-04741-2.pd

Datasets for transcriptomics, q-proteomics and phenotype microarrays of polyphosphate metabolism mutants from Escherichia coli

Indexación: Scopus.Author acknowledges Fondecyt Grants 1120209, 1121170 and Anillo ACT-1107Here, we provide the dataset associated with our research article on the polyphosphate metabolism entitled, “Multi-level evaluation of Escherichia coli polyphosphate related mutants using global transcriptomic, proteomic and phenomic analyses”. By integrating different omics levels (transcriptome, proteome and phenome), we were able to study Escherichia coli polyphosphate mutant strains (Δppk1, Δppx, and Δppk1-ppx). We have compiled here all datasets from DNA microarrys, q-proteomic (Isotope-Coded Protein Labeling, ICPL) and phenomic (Phenotype microarray) raw data we have obtained in all polyP metabolism mutants.http://www.sciencedirect.com/science/article/pii/S2352340917300860?via%3Dihu

Nitric oxide synthase-independent release of nitric oxide induced by KCl in the perfused mesenteric bed of the rat

The aim of the present study was to test whether the contractile responses elicited by KCl in the rat mesenteric bed are coupled to the release of nitric oxide (NO). Contractions induced by 70 mM KCl were coincident with the release of NO to the perfusate. The in vitro exposure to the nitric oxide synthase (NOS) inhibitor L-N(ω)-nitro-L-arginine methyl ester, L-NAME (1-100 μM) potentiated the vascular responses to 70 mM KCl and, unexpectedly, increased the KCl-stimulated release of NO. Moreover, even after the chronic treatment with L-NAME (70 mg/kg/day during 4 weeks), the KCl-induced release of NO was not reduced, whereas the potentiation of contractile responses was indeed achieved. The possibility that NOS had not been completely inhibited under our experimental conditions can be precluded because NOS activity was significantly inhibited after both L-NAME treatments. After the in vitro treatment with 1 to 100 μM L-NAME, the inhibition of NOS was concentration-dependent (from 50% to 90%). With regard to the basal release of NO, the inhibition caused by L-NAME was not concentration-dependent and reached a maximum of 40%, suggesting that basal NO outflow is only partially dependent on NOS activity. An eventual enhancement of NOS activity caused by KCl was disregarded because the activity of this enzyme measured in homogenates from mesenteric beds perfused with 70 mM KCl was significantly reduced. On the other hand, endothelium removal, employed as a negative control, almost abolished NOS activity, whereas the incubation with the Ca2+ ionophore A23187, employed as a positive control, induced an increase in NOS activity. It is concluded that in the mesenteric arterial bed of the rat, the contractile responses elicited by depolarization through KCl are coincident with a NOS-independent release of NO. This observation, which differs from the results obtained with noradrenaline, do not support the use of KCl as an alternative contractile agent whenever the participation of NO is under study. (C) 2000 Elsevier Science B.V.Fil: Mendizabal, Victoria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Farmacológicas. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Investigaciones Farmacológicas; ArgentinaFil: Poblete, I.. Pontificia Universidad Católica de Chile; ChileFil: Lomniczi, A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Besuhli, Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Huidobro Toro, J. P.. Pontificia Universidad Católica de Chile; ChileFil: Adler, Edda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Farmacológicas. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Investigaciones Farmacológicas; Argentin

La identidad en la encrucijada : migración peruana y educación en el Chile de hoy

La presente comunicación pretende poner de relieve los procesos sociales que han llevado a que durante el desarrollo de Chile como Estado Nación, haya cristalizado una cierta imagen de uniformidad social como forma de integración comunitaria, marginando en ello a diferentes grupos que se apartaban de los parámetros considerados de normalidad (principalmente los referentes indígenas y sectores populares de la vida política y social). A partir de estos procesos, surgirá una cierta idea de "identidad chilena", noción grupal cerrada, hegemónica y afiliada a caracteres nacionalistas, que se reproducirá socialmente a través de las diversas instituciones y discursos oficiales. En ese contexto, la escuela siempre fue utilizada como un instrumento útil para promover la anulación de las manifestaciones que no calzaban con los patrones ya establecidos, con aquello que se alejaba de la norma: maestros que sólo hablaran castellano en escuelas con presencia de niños indígenas, imitación de modelos europeos, entrega de contenidos que no guardaban relación con las particularidades del contexto o las culturas locales y una oferta curricular homogénea, fueron estrategias oficiales utilizadas prácticamente hasta la década del noventa del siglo pasado. La homogenización a través de la educación permitía así consolidar la construcción de un "ciudadano tipo" dentro del grupo social, excluyendo o normativizando otras autoidentificaciones que se alejaran de los propósitos de la construcción nacional. La "asimilación" pasó a ser la tónica de las diversas políticas públicas, configurando un discurso ideológico respecto de lo que se consideraba ser un/a "buen/a chileno/a". Este complejo escenario que se ha configurado en la construcción del estado nacional chileno a través del tiempo, es el que han encontrado algunos grupos migrados que han llegado a este país, especialmente peruanos, transformándose actualmente en la figura de subalternidad y alteridad por excelencia. Y si bien en diversos momentos de su historia social, el país impulsó políticas de inmigración, cabe destacar que siempre se caracterizaron, aunque no explícitamente, por una cierta priorización de origen nacional: los procesos de recepción de inmigrados europeos y las facilidades para adquirir los mismos derechos ciudadanos que cualquier nacional, resultan evidentes. El Estado no sólo ha actuado por razones económicas, sino que también ha planteado criterios de selección cultural e incluso a veces racial que han influido considerablemente en el conjunto de las relaciones interpersonales

Biochemistry, genetics and biotechnology of glycerol utilization in Pseudomonas species

Indexación: Scopus.The use of renewable waste feedstocks is an environment-friendly choice contributing to the reduction of waste treatment costs and increasing the economic value of industrial by-products. Glycerol (1,2,3-propanetriol), a simple polyol compound widely distributed in biological systems, constitutes a prime example of a relatively cheap and readily available substrate to be used in bioprocesses. Extensively exploited as an ingredient in the food and pharmaceutical industries, glycerol is also the main by-product of biodiesel production, which has resulted in a progressive drop in substrate price over the years. Consequently, glycerol has become an attractive substrate in biotechnology, and several chemical commodities currently produced from petroleum have been shown to be obtained from this polyol using whole-cell biocatalysts with both wild-type and engineered bacterial strains. Pseudomonas species, endowed with a versatile and rich metabolism, have been adopted for the conversion of glycerol into value-added products (ranging from simple molecules to structurally complex biopolymers, e.g. polyhydroxyalkanoates), and a number of metabolic engineering strategies have been deployed to increase the number of applications of glycerol as a cost-effective substrate. The unique genetic and metabolic features of glycerol-grown Pseudomonas are presented in this review, along with relevant examples of bioprocesses based on this substrate – and the synthetic biology and metabolic engineering strategies implemented in bacteria of this genus aimed at glycerol valorization.https://sfamjournals.onlinelibrary.wiley.com/doi/10.1111/1751-7915.1340

Editorial: Pathway, Genetic and Process Engineering of Microbes for Biopolymer Synthesis

Indexación ScopusTogether with the climate crisis, the heavy accumulation in oceans and soils of persistent pollutants, like polycyclic aromatics hydrocarbons and synthetic plastics, are the main drivers to impact nature and threaten human survival. It is particularly striking that most industrial polymers still originate from petrochemical sources—nearly 99% of the overall worldwide production. The result is materials that remain intact for centuries once deposited in the environment. Relying on non-renewable fossil chemicals limits our ability to establish a circular economy that promises to curb current emissions and contribute moderately to the global carbon cycle without surpassing its carrying capacity. For decades, commercial biopolymers have also been produced by microbial fermentation since nature has endowed many bacteria from urban sites to extreme environments (Orellana-Saez et al., 2019) with the enzymatic machinery to assemble these macromolecules. Despite the rapid pace of innovation, microbial biopolymers are still expensive to synthesize because the generally oxygen-intensive fermentation processes, downstream processing, and carbon feedstock cost boost production expenses (Oliveira et al., 2020). The biopolymers must additionally possess specific mechanical and physical properties to be processed industrially into products with a variety of applications (Moradali and Rehm, 2020).https://www.frontiersin.org/articles/10.3389/fbioe.2020.618383/ful

Factor V Leiden and prothrombin G20210A among Chilean patients with venous and arterial thrombosis

Iván Palomo. Departamento de Bioquímica Clínica e Inmunohematología, Facultad de Ciencias de la Salud. Universidad de Talca, Talca, Chile. Casilla 747,Talca, Chile.Factor V Leiden and G20210A mutation of prothrombin gene are two important genetic polymorphisms associated with an increased risk for thrombosis. Aim: To establish the prevalence of factor V Leiden and prothrombin G20210A mutation in the Chilean population and their association to venous and arterial thromboembolism. Material and methods: A case-control study was conducted where 149 patients with thrombosis (87 with arterial and 62 with venous thrombosis) confirmed by CAT-scan, electrocardiogram and cardiac enzymes or Doppler depending on the case, and 160 healthy blood donors were genetically analyzed for the presence of both polymorphisms. Results: Factor V Leiden mutation was found in 5.4% of patients and in 1.3% of healthy controls (p=0.04). Heterozygosity for G20210A prothrombin mutation was found in 5.4% of patients and in 2.5% of the control group (p=NS). When arterial and venous thrombosis were considered as separate entities, 4.6% of patients with arterial thrombosis and 6.5% with venous thrombosis presented factor V Leiden (p=NS). Likewise, 8.1% of patients with venous thrombosis and 3.5% of patients with arterial thrombosis had G20210A prothrombin mutation (p=NS). Conclusions: In non selected consecutive Chilean patients with arterial and venous thrombosis the frequency of factor V Leiden and prothrombin G20210A is less than we could expect from their prevalence in the general populatio

Oxidation of aldehydes used as food additives by peroxynitrite

Benzaldehyde and its derivatives are used as food supplements. These substances can be used mainly as flavorings or as antioxidants. Besides, peroxynitrite, an oxidizing agent, could be formed in canned food. Both species could react between them. The present article has focused on the kinetic study of the oxidation of aldehydes by peroxynitrite. A reaction mechanism that justifies all the experimental results is proposed. This mechanism, in acidic media, passes through three competitive pathways: (a) a radical attack that produces benzoic acid. (b) peracid oxidation, and (c) a nucleophilic attack of peroxynitrous acid over aldehyde to form an intermediate, X, that produces benzoic acid, or, through a Cannizzaro-type reaction, benzoic acid and benzyl alcohol. All rate constants involved in the third pathway (c) have been calculated. These results have never been described in the literature in acid media. A pH effect was analyzed

Fed-Batch mcl- Polyhydroxyalkanoates Production in Pseudomonas putida KT2440 and ΔphaZ Mutant on Biodiesel-Derived Crude Glycerol

Indexación ScopusCrude glycerol has emerged as a suitable feedstock for the biotechnological production of various industrial chemicals given its high surplus catalyzed by the biodiesel industry. Pseudomonas bacteria metabolize the polyol into several biopolymers, including alginate and medium-chain-length poly(3-hydroxyalkanoates) (mcl-PHAs). Although P. putida is a suited platform to derive these polyoxoesters from crude glycerol, the attained concentrations in batch and fed-batch cultures are still low. In this study, we employed P. putida KT2440 and the hyper-PHA producer ΔphaZ mutant in two different fed-batch modes to synthesize mcl-PHAs from raw glycerol. Initially, the cells grew in a batch phase (μmax 0.21 h–1) for 22 h followed by a carbon-limiting exponential feeding, where the specific growth rate was set at 0.1 (h–1), resulting in a cell dry weight (CDW) of nearly 50 (g L–1) at 40 h cultivation. During the PHA production stage, we supplied the substrate at a constant rate of 50 (g h–1), where the KT2440 and the ΔphaZ produced 9.7 and 12.7 gPHA L–1, respectively, after 60 h cultivation. We next evaluated the PHA production ability of the P. putida strains using a DO-stat approach under nitrogen depletion. Citric acid was the main by-product secreted by the cells, accumulating in the culture broth up to 48 (g L–1) under nitrogen limitation. The mutant ΔphaZ amassed 38.9% of the CDW as mcl-PHA and exhibited a specific PHA volumetric productivity of 0.34 (g L–1 h–1), 48% higher than the parental KT2440 under the same growth conditions. The biosynthesized mcl-PHAs had average molecular weights ranging from 460 to 505 KDa and a polydispersity index (PDI) of 2.4–2.6. Here, we demonstrated that the DO-stat feeding approach in high cell density cultures enables the high yield production of mcl-PHA in P. putida strains using the industrial crude glycerol, where the fed-batch process selection is essential to exploit the superior biopolymer production hallmarks of engineered bacterial strains. © Copyright © 2021 Borrero-de Acuña, Rohde, Saldias and Poblete-Castro.https://www.frontiersin.org/articles/10.3389/fbioe.2021.642023/ful

Intraspecific variability of popcorn S7 lines for phosphorus efficiency in the soil.

The expansion of agriculture, coupled with the need for sustainable cropping, is one of the greatest challenges of the scientific community working on the generation of new cultivars adapted to abiotic stress conditions. The aim of this study was to evaluate the variability of popcorn lines as to responsiveness and efficiency in phosphorus use, as a first step towards the implementation of a breeding program interested in the practice of sustainable agriculture. Twenty-five popcorn lines were evaluated in two locations with different phosphorus levels in the soil, using a randomized block design. The following traits were measured: plant height, ear height, female flowering date, male flowering date, male-female flowering interval, ear diameter, ear length, 100-grain weight, grain yield, popping expansion, and expanded popcorn volume per hectare. A combined analysis of variance and test of means were performed, and the lines were classified as to their phosphorus use efficiency, according to their production performance in the different environments. The genetic diversity between the lines was estimated by Tocher?s and UPGMA clustering methods, using generalized Mahalanobis distance. Lines L59, P7, P2, P3, P4, P8, P10, P9, L66, L70, L69, and P5 were efficient and responsive, whereas lines L75, L80, L61, L77, L63, L65, P1, L54, L53, L88, and L71 were inefficient and nonresponsive. Genetic variability was greater in the environments with low phosphorus in the soil, suggesting that the selection pressure exerted in the stressing environment is a decisive factor to obtain a higher expression of variability