41 research outputs found
Aminopeptidase de Mesorhizobium sp. descoberta por mineração de dados genômicos com aplicações biotecnológicas e seu impacto na fisiologia bacteriana
No full textA análise de mineração de dados genômicos de Mesorhizobium sp. revelou a presença de uma ORF de 1257pb contendo o gene mesoamp, que codifica uma proteína de 418 aminoácidos. A sequência de aminoácidos deduzida apresenta 50% de identidade com uma amino-peptidase termoestável de Thermus thermophilus, membro da família de peptidase M29, e oito assinaturas caraterísticas das metalo-protease termofílicas foram encontrados. O gene mesoamp foi clonado e expresso em Escherichia coli. A massa molecular da proteína foi avaliada por SDS-PAGE e filtração em gel, o que indicou que a proteína apresenta 45,72kDa e 88,05kDa, respectivamente, sugerindo uma estrutura dimérica da enzima recombinante. A enzima foi nomeada como MesoAmp. Em seguida realizou-se a modelagem 3D estrutural, que mostrou uma região de dimerização e uma região de ligação ao substrato altamente conservada contendo os resíduos de ligação a metais e o resíduo catalítico. A enzima apresentou atividade ótima em pH 8,5 e temperatura de 45°C, foi fortemente ativada por Co2+ e Mn2+, nestas mesmas condições de reação, a enzima apresentou Km e Kcat de 0,2364±0,018mM e 712,1±88,12seg-1, respectivamente. Além disso, foi verificado uma notável estabilidade da enzima em solventes orgânicos e elevadas concentrações de NaCl, além de um alto grau de reutilização sem perda apreciável de atividade o que torna MesoAmp única, este trabalho estabelece as bases para potenciais aplicações biotecnológicas e/ou o desenvolvimento de tecnologias sustentáveis, além de descrever uma das primeiras aminopeptidases solvente e halo-tolerantes identificados para o gênero Mesorhizobium sp.The genomic data mining analysis of a Mesorhizobium sp. revealed the presence of a 1257-bp open reading frame containing the Mesoamp gene, which encodes a protein of 418 amino acids. The deduced amino acid sequence was 50% identical to the thermostable amino-peptidase from Thermus thermophilus, a member of peptidase family M29, and eight fingerprints signatures for a thermophilic metalloprotease were found. The mesoamp gene was cloned and overexpressed in Escherichia coli. The molecular mass of the protein was assessed by SDS-PAGE and gel filtration, which indicated the protein weighs 45.72kDa and 88.05kDa, respectively, suggesting a dimeric structure of the recombinant enzyme. The enzyme was designated as MesoAmp. The 3D structural modeling showed a dimerization region with highly conserved catalytic and binding metal residues. The enzyme exhibited optimum activity at pH 8.5 and 45°C and was strongly activated by Co2+ and Mn2+ . Under these reaction conditions, the enzyme displayed Km and Kcat values of 0.2364±0.018mM and 712.1±88.12seg-1 , respectively. Additionally, the remarkable stability of the enzyme in organic solvents, its activity at high concentrations of NaCl and the high degree of reuse without appreciable loss of activity makes MesoAmp unique. In summary, this work lays the foundation for potential biotechnological applications and/or the development of environmentally friendly technologies and describes the first solvent and halo-tolerant amino-peptidases identified for the Mesorhizobium sp genus.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES
Aminopeptidase de Mesorhizobium sp. descoberta por mineração de dados genômicos com aplicações biotecnológicas e seu impacto na fisiologia bacteriana
No full textA análise de mineração de dados genômicos de Mesorhizobium sp. revelou a presença de uma ORF de 1257pb contendo o gene mesoamp, que codifica uma proteína de 418 aminoácidos. A sequência de aminoácidos deduzida apresenta 50% de identidade com uma amino-peptidase termoestável de Thermus thermophilus, membro da família de peptidase M29, e oito assinaturas caraterísticas das metalo-protease termofílicas foram encontrados. O gene mesoamp foi clonado e expresso em Escherichia coli. A massa molecular da proteína foi avaliada por SDS-PAGE e filtração em gel, o que indicou que a proteína apresenta 45,72kDa e 88,05kDa, respectivamente, sugerindo uma estrutura dimérica da enzima recombinante. A enzima foi nomeada como MesoAmp. Em seguida realizou-se a modelagem 3D estrutural, que mostrou uma região de dimerização e uma região de ligação ao substrato altamente conservada contendo os resíduos de ligação a metais e o resíduo catalítico. A enzima apresentou atividade ótima em pH 8,5 e temperatura de 45°C, foi fortemente ativada por Co2+ e Mn2+, nestas mesmas condições de reação, a enzima apresentou Km e Kcat de 0,2364±0,018mM e 712,1±88,12seg-1, respectivamente. Além disso, foi verificado uma notável estabilidade da enzima em solventes orgânicos e elevadas concentrações de NaCl, além de um alto grau de reutilização sem perda apreciável de atividade o que torna MesoAmp única, este trabalho estabelece as bases para potenciais aplicações biotecnológicas e/ou o desenvolvimento de tecnologias sustentáveis, além de descrever uma das primeiras aminopeptidases solvente e halo-tolerantes identificados para o gênero Mesorhizobium sp.The genomic data mining analysis of a Mesorhizobium sp. revealed the presence of a 1257-bp open reading frame containing the Mesoamp gene, which encodes a protein of 418 amino acids. The deduced amino acid sequence was 50% identical to the thermostable amino-peptidase from Thermus thermophilus, a member of peptidase family M29, and eight fingerprints signatures for a thermophilic metalloprotease were found. The mesoamp gene was cloned and overexpressed in Escherichia coli. The molecular mass of the protein was assessed by SDS-PAGE and gel filtration, which indicated the protein weighs 45.72kDa and 88.05kDa, respectively, suggesting a dimeric structure of the recombinant enzyme. The enzyme was designated as MesoAmp. The 3D structural modeling showed a dimerization region with highly conserved catalytic and binding metal residues. The enzyme exhibited optimum activity at pH 8.5 and 45°C and was strongly activated by Co2+ and Mn2+ . Under these reaction conditions, the enzyme displayed Km and Kcat values of 0.2364±0.018mM and 712.1±88.12seg-1 , respectively. Additionally, the remarkable stability of the enzyme in organic solvents, its activity at high concentrations of NaCl and the high degree of reuse without appreciable loss of activity makes MesoAmp unique. In summary, this work lays the foundation for potential biotechnological applications and/or the development of environmentally friendly technologies and describes the first solvent and halo-tolerant amino-peptidases identified for the Mesorhizobium sp genus.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES
Aminopeptidase from a Mesorhizobium sp. discovered by genomic data mining with potential biotechnology application and its impact on bacterial physiology
No full textA análise de mineração de dados genômicos de Mesorhizobium sp. revelou a presença de uma ORF de 1257pb contendo o gene mesoamp, que codifica uma proteína de 418 aminoácidos. A sequência de aminoácidos deduzida apresenta 50% de identidade com uma amino-peptidase termoestável de Thermus thermophilus, membro da família de peptidase M29, e oito assinaturas caraterísticas das metalo-protease termofílicas foram encontrados. O gene mesoamp foi clonado e expresso em Escherichia coli. A massa molecular da proteína foi avaliada por SDS-PAGE e filtração em gel, o que indicou que a proteína apresenta 45,72kDa e 88,05kDa, respectivamente, sugerindo uma estrutura dimérica da enzima recombinante. A enzima foi nomeada como MesoAmp. Em seguida realizou-se a modelagem 3D estrutural, que mostrou uma região de dimerização e uma região de ligação ao substrato altamente conservada contendo os resíduos de ligação a metais e o resíduo catalítico. A enzima apresentou atividade ótima em pH 8,5 e temperatura de 45°C, foi fortemente ativada por Co2+ e Mn2+, nestas mesmas condições de reação, a enzima apresentou Km e Kcat de 0,2364±0,018mM e 712,1±88,12seg-1, respectivamente. Além disso, foi verificado uma notável estabilidade da enzima em solventes orgânicos e elevadas concentrações de NaCl, além de um alto grau de reutilização sem perda apreciável de atividade o que torna MesoAmp única, este trabalho estabelece as bases para potenciais aplicações biotecnológicas e/ou o desenvolvimento de tecnologias sustentáveis, além de descrever uma das primeiras aminopeptidases solvente e halo-tolerantes identificados para o gênero Mesorhizobium sp.The genomic data mining analysis of a Mesorhizobium sp. revealed the presence of a 1257-bp open reading frame containing the Mesoamp gene, which encodes a protein of 418 amino acids. The deduced amino acid sequence was 50% identical to the thermostable amino-peptidase from Thermus thermophilus, a member of peptidase family M29, and eight fingerprints signatures for a thermophilic metalloprotease were found. The mesoamp gene was cloned and overexpressed in Escherichia coli. The molecular mass of the protein was assessed by SDS-PAGE and gel filtration, which indicated the protein weighs 45.72kDa and 88.05kDa, respectively, suggesting a dimeric structure of the recombinant enzyme. The enzyme was designated as MesoAmp. The 3D structural modeling showed a dimerization region with highly conserved catalytic and binding metal residues. The enzyme exhibited optimum activity at pH 8.5 and 45°C and was strongly activated by Co2+ and Mn2+ . Under these reaction conditions, the enzyme displayed Km and Kcat values of 0.2364±0.018mM and 712.1±88.12seg-1 , respectively. Additionally, the remarkable stability of the enzyme in organic solvents, its activity at high concentrations of NaCl and the high degree of reuse without appreciable loss of activity makes MesoAmp unique. In summary, this work lays the foundation for potential biotechnological applications and/or the development of environmentally friendly technologies and describes the first solvent and halo-tolerant amino-peptidases identified for the Mesorhizobium sp genus.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES
Principales herramientas moleculares empleadas en la ciencia animal
No full textLa domesticación animal y la ganadería son actividades desarrolladas por el hombre desde hace unos 10000 años, que ha permitido obtener una asombrosa cantidad de razas y atributos en especies animales, que han cubierto parte de las necesidades alimentarias, utilitarias y culturales del hombre. Con el desarrollo de la genética y la biología molecular durante el siglo XX, se han logrado optimizar muchas de esas prácticas milenarias. El progreso de técnicas como la PCR, la detección de marcadores moleculares, la secuenciación de DNA, RNA y proteínas, así como las herramientas bioinformáticas, han permitido conocer la constitución y organización del genoma de numerosas especies, su variación, la relación fenotipo-genotipo, la expresión genética y el metabolismo. Asimismo, con estos avances se han perfeccionado técnicas en las ciencias pecuarias, como el diagnóstico de enfermedades y zoonosis, la selección asistida, el mejoramiento genético, la reproducción, el estudio de diversidad genética y nutrición animal y el desarrollo de tecnologías de producción. Aun cuando muchas de los métodos de manejo y mejoramiento tradicionales no podrán ser reemplazadas, es necesaria la inclusión de nuevas técnicas moleculares, por lo tanto la presente revisión describe las principales tecnologías aplicadas en esta área, demostrando como el uso de herramientas moleculares es cada vez más necesario en términos técnicos y económicos.</jats:p
Principales herramientas moleculares empleadas en la ciencia animal
No full textThe animal domestication and livestock activities are developed by man since about 10,000 years ago, which has led to a staggering number of animal breeds and attributes, which have covered part of the need food, cultural and utility of man. With development of genetics and molecular biology in the XX century, has been made to optimized many of these ancient practices. The progress of techniques such as PCR, detection of molecular markers, sequencing of DNA, RNA and proteins, and bioinformatics tools, have allowed to know the constitution and organization of the genome of many species, its variation, genotype-phenotype relationship, gene expression and metabolism. In addition, these advances have perfected techniques in animal science, such as diagnosis of diseases and zoonoses, assisted selection, breeding, reproduction, the study of genetic diversity, animal nutrition and the development of production technologies. Although many of the methods of management and traditional breeding may not be replaced, it is necessary to include new molecular techniques, therefore the present review describes the main technologies applied in this area, demonstrating how the use of molecular tools is increasingly more necessary in technical and economic terms.La domesticación animal y la ganadería son actividades desarrolladas por el hombre desde hace unos 10000 años, que ha permitido obtener una asombrosa cantidad de razas y atributos en especies animales, que han cubierto parte de las necesidades alimentarias, utilitarias y culturales del hombre. Con el desarrollo de la genética y la biología molecular durante el siglo XX, se han logrado optimizar muchas de esas prácticas milenarias. El progreso de técnicas como la PCR, la detección de marcadores moleculares, la secuenciación de DNA, RNA y proteínas, así como las herramientas bioinformáticas, han permitido conocer la constitución y organización del genoma de numerosas especies, su variación, la relación fenotipo-genotipo, la expresión genética y el metabolismo. Asimismo, con estos avances se han perfeccionado técnicas en las ciencias pecuarias, como el diagnóstico de enfermedades y zoonosis, la selección asistida, el mejoramiento genético, la reproducción, el estudio de diversidad genética y nutrición animal y el desarrollo de tecnologías de producción. Aun cuando muchas de los métodos de manejo y mejoramiento tradicionales no podrán ser reemplazadas, es necesaria la inclusión de nuevas técnicas moleculares, por lo tanto la presente revisión describe las principales tecnologías aplicadas en esta área, demostrando como el uso de herramientas moleculares es cada vez más necesario en términos técnicos y económicos
Principales herramientas moleculares empleadas en la ciencia animal
No full textThe animal domestication and livestock activities are developed by man since about 10,000 years ago, which has led to a staggering number of animal breeds and attributes, which have covered part of the need food, cultural and utility of man. With development of genetics and molecular biology in the XX century, has been made to optimized many of these ancient practices. The progress of techniques such as PCR, detection of molecular markers, sequencing of DNA, RNA and proteins, and bioinformatics tools, have allowed to know the constitution and organization of the genome of many species, its variation, genotype-phenotype relationship, gene expression and metabolism. In addition, these advances have perfected techniques in animal science, such as diagnosis of diseases and zoonoses, assisted selection, breeding, reproduction, the study of genetic diversity, animal nutrition and the development of production technologies. Although many of the methods of management and traditional breeding may not be replaced, it is necessary to include new molecular techniques, therefore the present review describes the main technologies applied in this area, demonstrating how the use of molecular tools is increasingly more necessary in technical and economic terms.La domesticación animal y la ganadería son actividades desarrolladas por el hombre desde hace unos 10000 años, que ha permitido obtener una asombrosa cantidad de razas y atributos en especies animales, que han cubierto parte de las necesidades alimentarias, utilitarias y culturales del hombre. Con el desarrollo de la genética y la biología molecular durante el siglo XX, se han logrado optimizar muchas de esas prácticas milenarias. El progreso de técnicas como la PCR, la detección de marcadores moleculares, la secuenciación de DNA, RNA y proteínas, así como las herramientas bioinformáticas, han permitido conocer la constitución y organización del genoma de numerosas especies, su variación, la relación fenotipo-genotipo, la expresión genética y el metabolismo. Asimismo, con estos avances se han perfeccionado técnicas en las ciencias pecuarias, como el diagnóstico de enfermedades y zoonosis, la selección asistida, el mejoramiento genético, la reproducción, el estudio de diversidad genética y nutrición animal y el desarrollo de tecnologías de producción. Aun cuando muchas de los métodos de manejo y mejoramiento tradicionales no podrán ser reemplazadas, es necesaria la inclusión de nuevas técnicas moleculares, por lo tanto la presente revisión describe las principales tecnologías aplicadas en esta área, demostrando como el uso de herramientas moleculares es cada vez más necesario en términos técnicos y económicos
Aislamiento, selección y caracterización de cepas del género lactobacillus aisladas de líquido ruminal vacuno en la zona Sur del Lago, Venezuela
No full textLactic acid bacteria of Lactobacillus genus were isolated from cattle rumen fluid in the Colon, was carried out by sampling animals post-mortem at the at slaughter and distribution service meat “FIBASA”. These samples were diluted to different concentrations ranging from 10-1 - 10-7 and plated on, thereby obtaining bacterial colonies, which were evaluated in vitro with growth tests, shape, color, size, mobility, presence of spores, lactic acid production, fermentation of carbohydrates, acid resistance (2.5 to 6.5) and temperature profiles from (4 °C - 50 °C). This was achieved characterize two strains (L1 and L2), whose biochemical and morphological analysis places them within the genus Lactobacillus, These strains have high antagonistic activity against pathogenic bacteria which makes them a probiotic potential, for animal feed. La caracterización de bacterias ácido lácticas del género Lactobacillus aisladas y seleccionadas de líquido ruminal vacuno en el Municipio Colon del estado Zulia, se llevó a cabo mediante el muestreo de animales post-morten en el centro de matanza y servicio de distribución de carnes “FIBASA”. Dichas muestras fueron diluidas a diferentes concentraciones desde 10-1 – 10-7 y sembradas en placas, obteniendo con ello colonias bacterianas, las cuales fueron evaluadas in-vitro con pruebas de crecimiento, forma, color, tamaño, movilidad, presencia de esporas, producción de ácido láctico, fermentación de carbohidratos, resistencia a ácidos (2,5 – 6,5) y perfiles térmicos desde (4°C – 50°C). Con ello se logró caracterizar dos cepas (L1 y L2), cuyo análisis bioquímico y morfológico las ubica dentro del género Lactobacillus, estas cepas presentaron una alta actividad antagónica contra bacterias patógenas, los que las convierte en un potencial probiótico para la alimentación animal
Legal Protection Application of Victims Through a Combined Lawsuit for Compensation in Case of Criminal Acts of Fraud and Money Laundering (Case Study: PT First Travel Cassation Decision Number 3096 K/Pid.Sus/2018)
No full textVictims in the criminal justice system are forgotten and disadvantaged subjects. In addition to the victim having suffered losses as a result of the crime that occurred to him, both materially, physically and psychologically, the victim must also suffer because unknowingly they are often treated only as a means for the realization of legal certainty. As happened in the case of fraud and money laundering by PT First Travel. In the cassation decision in the PT First Travel case, the judge upheld the decision at the first level and at the appellate level. This decision resulted in victims of prospective first travel pilgrims not receiving compensation because the first travel assets were confiscated and returned to the state, even though the state was not harmed at all in this case. Therefore the author is interested in conducting research on cases of fraud and money laundering by PT First Travel about how victims of first travel can get their rights back through a combination of claims for compensation because the Criminal Procedure Code has provided a way for victims to be able to get their rights in Article 98 -101 KUHAP. The author is interested in discussing 1) How important is the combined lawsuit for compensation in cases of fraud and money laundering at PT First Travel in the context of victim protection? 2) How is the cassation decision Number 3096 K/Pid.Sus/2018 reviewed based on the theory of justice and the theory of expediency?. This legal research is normative research using library materials or secondary data. Based on the results of the research that the authors have done, the results show that: 1) The importance of combined lawsuit for compensation in cases of fraud and money laundering at PT Frst Travel is as a way to provide protection for victims so they can get their rights back. Even though compensation is in the realm of civil law, the Criminal Procedure Code has provided a way through a positive relationship in Article 98 of the Criminal Procedure Code which allows cases for claims for compensation to be combined with criminal cases at the same time. The reason for not carrying out the merger in the first travel case was due to the absence of a request from the victim, this could have happened due to the victim's ignorance of the existence of Article 98 of the Criminal Procedure Code. the theory of justice and expediency, this can be seen from the side of the victim who has been harmed but does not get his rights in the form of compensation. Of course the cassation decision did not provide justice and benefits for the victim
Caracterización fenotípica y genotípica de cultivares de cacao (Theobroma cacao L.) de Dibulla, La Guajira, Colombia
No full textIn Sierra Nevada de Santa Marta, cacao plantations are comprised of commercial hybrid cultivars, and although native cacaos are found, they are not widely cultivated. Given the need to verify if these cacao varieties found in the Sierra region belong to the Criollo type genetic group, a phenotypic and genotypic characterization of cacao from the municipality Dibulla, La Guajira was carried out. For this, 11 cultivars were sampled in Mingueo. Phenotypic traits were evaluated using UPOV descriptors for cacao. The qualitative and quantitative parameters were compared through cluster and principal component analyses (PCA), and the quantitative variables through the non-parametric Mann-Whitney test. Molecular biology protocols were standardized, and the ITS region was sequenced to assess genetic relationships. From the sequences, groupings were carried out utilizing distance and phylogenetic methods. Finally, significant differences were found among the seeds (p = 0.01), and the white coloration of the cotyledon of the criollos or native stands out in contrast to the dark purple coloration of the hybrids. The cluster analysis, PCA, and sequence analysis groupings, showed differences between the group of native cacaos and commercial hybrids cultivated; in addition, native cacaos are related to the Criollo type group.En la Sierra Nevada de Santa Marta los cultivos de cacao están conformados mayoritariamente por cultivares híbridos comerciales y, aunque se encuentran cacaos nativos, estos son poco cultivados. Dada la necesidad de verificar si estos cultivares de cacao encontrados en la Sierra pertenecen al grupo genético tipo Criollo, se realizó una caracterización fenotípica y genotípica de cacaos del municipio Dibulla, La Guajira. Para esto, se muestrearon 11 cultivares en Mingueo. Los rasgos fenotípicos se evaluaron empleando descriptores UPOV para cacao. Los parámetros cualitativos y cuantitativos se cotejaron por análisis de conglomerado y análisis de componentes principales (ACP), y las variables cuantitativas se compararon a través de la prueba no paramétrica test de Mann-Whitney. Para evaluar las relaciones genéticas, se estandarizaron protocolos de biología molecular y se secuenció la región ITS. A partir de las secuencias, se realizaron agrupamientos por métodos de distancia y filogenéticos. Finalmente, se encontraron diferencias significativas entre las semillas (p = 0,01), y resalta la coloración blanca del cotiledón de los criollos en contraste con la coloración púrpura oscura de los híbridos. Asimismo, los análisis de conglomerados, ACP y los análisis de secuencias demostraron diferencias entre el grupo de los cacaos nativos y los híbridos comerciales cultivados; además, los cacaos nativos se emparentan con el grupo de cacao tipo Criollo
Aislamiento, selección y caracterización de cepas del género lactobacillus aisladas de líquido ruminal vacuno en la zona Sur del Lago, Venezuela
No full textLactic acid bacteria of Lactobacillus genus were isolated from cattle rumen fluid in the Colon, was carried out by sampling animals post-mortem at the at slaughter and distribution service meat “FIBASA”. These samples were diluted to different concentrations ranging from 10-1 - 10-7 and plated on, thereby obtaining bacterial colonies, which were evaluated in vitro with growth tests, shape, color, size, mobility, presence of spores, lactic acid production, fermentation of carbohydrates, acid resistance (2.5 to 6.5) and temperature profiles from (4 °C - 50 °C). This was achieved characterize two strains (L1 and L2), whose biochemical and morphological analysis places them within the genus Lactobacillus, These strains have high antagonistic activity against pathogenic bacteria which makes them a probiotic potential, for animal feed. La caracterización de bacterias ácido lácticas del género Lactobacillus aisladas y seleccionadas de líquido ruminal vacuno en el Municipio Colon del estado Zulia, se llevó a cabo mediante el muestreo de animales post-morten en el centro de matanza y servicio de distribución de carnes “FIBASA”. Dichas muestras fueron diluidas a diferentes concentraciones desde 10-1 – 10-7 y sembradas en placas, obteniendo con ello colonias bacterianas, las cuales fueron evaluadas in-vitro con pruebas de crecimiento, forma, color, tamaño, movilidad, presencia de esporas, producción de ácido láctico, fermentación de carbohidratos, resistencia a ácidos (2,5 – 6,5) y perfiles térmicos desde (4°C – 50°C). Con ello se logró caracterizar dos cepas (L1 y L2), cuyo análisis bioquímico y morfológico las ubica dentro del género Lactobacillus, estas cepas presentaron una alta actividad antagónica contra bacterias patógenas, los que las convierte en un potencial probiótico para la alimentación animal
