Low-Scale Expression and Purification of an Active Putative Iduronate 2-Sulfate Sulfatase-Like Enzyme from Escherichia coli K12

The sulfatase family involves a group of enzymes with a large degree of similarity. Until now, sixteen human sulfatases have been identified, most of them found in lysosomes. Human deficiency of sulfatases generates various genetic disorders characterized by abnormal accumulation of sulfated intermediate compounds. Mucopolysaccharidosis type II is characterized by the deficiency of iduronate 2-sulfate sulfatase (IDS), causing the lysosomal accumulation of heparan and dermatan sulfates. Currently, there are several cases of genetic diseases treated with enzyme replacement therapy, which have generated a great interest in the development of systems for recombinant protein expression. In this work we expressed the human recombinant IDS-Like enzyme (hrIDS-Like) in Escherichia coli DH5α. The enzyme concentration revealed by ELISA varied from 78. 13 to 94. 35 ng/ml and the specific activity varied from 34. 20 to 25. 97 nmol/h/mg. Western blotting done after affinity chromatography purification showed a single band of approximately 40 kDa, which was recognized by an IgY polyclonal antibody that was developed against the specific peptide of the native protein. Our 100 ml-shake-flask assays allowed us to improve the enzyme activity seven fold, compared to the E. coli JM109/pUC13-hrIDS-Like system. Additionally, the results obtained in the present study were equal to those obtained with the Pichia pastoris GS1115/pPIC-9-hrIDS-Like system (3 L bioreactor scale). The system used in this work (E. coli DH5α/pGEX-3X-hrIDS-Like) emerges as a strategy for improving protein expression and purification, aimed at recombinant protein chemical characterization, future laboratory assays for enzyme replacement therapy, and as new evidence of active putative sulfatase production in E. coli. © 2013 The Microbiological Society of Korea and Springer-Verlag Berlin Heidelberg.Fil: Morales Álvarez, Edwin David. Universidad del Quindio; ColombiaFil: Rivera Hoyos, Claudia Marcela. Universidad del Quindio. Facultad de Medicina; ColombiaFil: Baena Moncada, Angelica Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Landázuri, Patricia. Universidad del Quindio. Facultad de Medicina. Centro de Investig. Biomédicas; ColombiaFil: Poutou Piñales, Raúl A.. Pontificia Universidad Javeriana; ColombiaFil: Sáenz Suárez, Homero. Universidad del Quindio; ColombiaFil: Barrera, Luis A.. Pontificia Universidad Javeriana; ColombiaFil: Echeverri Peña, Olga Y.. Pontificia Universidad Javeriana; Colombi

Inducción a la formación profesional

Desarrolla a manera de módulos, la forma y la información que de se debe entregar a los aprendices al momento de que estos ingresen a la institución para conocer el contexto, deberes, derechos, entre otros, que los estarán rodeando en el ámbito académico.It develops as a module, the form and the information that must be given to the apprentices when they enter the institution to know the context, duties, rights, among others, that will be surrounding them in the academic field.Conocimiento e Integración Grupal -- Inducción al SENA -- Inducción a la Formación Profesional Integral -- Inducción al Centro, Programa o Proyecto -- Inducción al Programa de Formación Profesional -- Inducción a los Servicios al Alumnado -- Servicios al Alumnado: Biblioteca -- Servicios al Alumnado: Promoción y contratación de Aprendices -- Servicios al Alumnado: Trabajo Social -- Servicios al Alumnado: Capellanía -- Servicios al Alumnado: Registro y Certificación -- Servicios al Alumnado: Validación -- Servicios al Alumnado: Actividades Recreativas, Culturales y Deportivas.naPara sujetos de formación40 página

1H-nuclear magnetic resonance analysis of urine as diagnostic tool for organic acidemias and aminoacidopathies

The utility of low-resolution H-NMR analysis for the identification of biomarkers provided evidence for rapid biochemical diagnoses of organic acidemia and aminoacidopathy. H-NMR, with a sensitivity expected for a field strength of 400 MHz at 64 scans was used to establish the metabolomic urine sample profiles of an infant population diagnosed with small molecule Inborn Errors of Metabolism (smIEM) compared to unaffected individuals. A qualitative differentiation of the H-NMR spectral profiles of urine samples obtained from individuals affected by different organic acidemias and aminoacidopathies was achieved in combination with GC-MS. The smIEM disorders investigated in this study included phenylalanine metabolism; isovaleric, propionic, 3-methylglutaconicm and glutaric type I acidemia; and deficiencies in medium chain acyl-coenzyme and holocarboxylase synthase. The observed metabolites were comparable and similar to those reported in the literature, as well as to those detected with higher-resolution NMR. In this study, diagnostic marker metabolites were identified for the smIEM disorders. In some cases, changes in metabolite profiles differentiated post-treatments and follow-ups while allowing for the establishment of different clinical states of a biochemical disorder. In addition, for the first time, a H-NMR-based biomarker profile was established for holocarboxylase synthase deficiency spectrum

Research, diagnosis and education in inborn errors of metabolism in Colombia: 20 years’ experience from a reference center

Abstract The use of specialized centers has been the main alternative for an appropriate diagnosis, management and follow up of patients affected by inborn errors of metabolism (IEM). These centers facilitate the training of different professionals, as well as the research at basic, translational and clinical levels. Nevertheless, few reports have described the experience of these centers and their local and/or global impact in the study of IEM. In this paper, we describe the experience of a Colombian reference center for the research, diagnosis, training and education on IEM. During the last 20 years, important advances have been achieved in the clinical knowledge of these disorders, as well as in the local availability of several diagnosis tests. Organic acidurias have been the most frequently detected diseases, followed by aminoacidopathies and peroxisomal disorders. Research efforts have been focused in the production of recombinant proteins in microorganisms towards the development of new enzyme replacement therapies, the design of gene therapy vectors and the use of bioinformatics tools for the understanding of IEM. In addition, this center has participated in the education and training of a large number professionals at different levels, which has contributed to increase the knowledge and divulgation of these disorders along the country. Noteworthy, in close collaboration with patient advocacy groups, we have participated in the discussion and construction of initiatives for the inclusion of diagnosis tests and treatments in the health system

Evaluación de la ecofisiología digestiva y sus aplicaciones biotecnológicas en bovinos criollos BON

IP 1118-07-106-96v.1. Informe Final. -- v.2. Determinacion de las concentracionesde amoniaco en liquido ruminal y urea serica en tres razas bovinas (bon, cebu, holstein) alimentadoscon dos calidades de forraje / Liliana Maria Osorio Muñoz, Jaime Alberto Orrego Bustamante. -- v.3. Cuantificacion deagv y ph en el liquido ruminal de tres razas de bovinos (bon, holstein y cebu) alimentados con doscalidadesde forraje / Monica Maria Gil Espinosa, Silvia Margarita Quiros Gomez. -- v.4. Resultados deInvestigacion.Quiros, Orlando Ruiz. -- En: Revista Colombiana de Ciencias Pecuarias. --Vol. 12 (noviembre 11 y 12 de 1999);p. 61. -- Estandarizacion de las tecnicas de microscopia de luzy electronica para determinar la colonizacion;de los h.a.r a tejidos vegetales del pasto para (brachiaria mutica) / Alfonso Giraldo, I.Cano. -- En: Revista;Colombiana de Ciencias Pecuarias. -- Vol. 12 (noviembre 11y 12de 1999);p. 129. -- Aplicacion de la tecnica;de microscopia electronica de barrido (meb) para determinar lacolonizacion de los h.a.r a los tejidos;vegetales del pasto para (brachiaria mutica) / Alfonso Giraldo,I. Cano, D. Polanco. -- En: Revista Colombiana;de Ciencias Pecuarias. -- Vol. 12 (noviembre 11 y 12 de 1999);p. 129. --Estandarizacion de la tecnica de;mordantar fibra con cromio como marcador externo, para estimarvarios parametros de la cinetica digestiva en;rumiantes / Silvia Quiros, Alfonso Giraldo. -- En: Revista Colombiana deCiencias Pecuarias. -- No. 3;(diciembre de 1998); p. 124. -- Degradabilidad ruminal insitudel forrajede alta y baja calidad de pasto;kikuyo a diferentes tiempos de incubacion y en tres razasde bovinos / Alfonso Giraldo, Mauricio Agudelo, Juan;Munera. -- En: Revista Colombiana de Ciencias Pecuarias. -'- no.4 (mayo 1999); p. 2. -- ISSN: 0120-0690.;ARTICULO(S) EN REVISTA: Pastos y forrajes, reproduccion yproduccion animal : estandarizacion de la tecnica;de la biodegradacion ruminal in situ, para evaluar forrajes tropicales. --En: Revista Colombiana de Ciencias;Pecuarias. -- Vol. 9 (diciembre 1996); p. 59-63. -- Proyecto regional : evaluacion de la ecofisiologia;mediante la utilizacion de tres tecnicas de medicion de ladigestibilidad/ Giraldo Luis Alfonso, Luis G;Echeverri, Alejandro Velez. -- En: Revista Colombiana de Ciencias Pecuarias. -- Vol. 12 (noviembre 11 y 12 de;1999); p. 47. -- Biodegradabilidad ruminal in situ en doscalidades de pasto estrella (cynodon nlemfuensis) /;Luis Alfonso Giraldo, R. Galvis, J. C. Carmona, S. Ayala.'-- en:Revista Colombiana de Ciencias Pecuarias. --;Vol. 12 (noviembre 11 y 12 de 1999); p. 55. -- Cuantificaciondeagv ph enliquido ruminal de tres razas de;bovinos (bon, holstein y cebu) alimentados con dos clasesde forraje / Alfonso Giraldo, Monica Gil E., Silvia;(noviembre 11 y 12 de 1999); p. 41. -- Estimacion del consumodeforraje de pasto para (b. Mutica) en cabros,;digestiva y sus aplicaciones biotecnicas en bovinos criollos bon/ GiraldoLuis A., Cano Ida Y. -- En: Revista;Colombiana de Ciencias Pecuarias. -- Vol. 10, no. 317 (noviembre6 y 7 de1997); p. 1-3. -- ISSN 01200690. --;Degradabilidad ruminal in situ del forraje de alta y bajacalidad de pastokikuyo en tres razas de bovinos /;Giraldo Alfonso, Agudelo Mauricio, Munera Juan. -- En: RevistaColombianade Ciencias Pecuarias. -- Vol. 1