Abstract - Human sulfatase transiently and functionally active expressed in E. coli K12

Human sulfatase transiently and functionally active expressed in E. coli K12

Raúl A. Poutou Piñales
Instituto de Errores Innatos del Metabolismo
Grupo de Biotecnología Ambiental e Industrial
Facultad de Ciencias
Pontificia Universidad Javeriana
Cra. 7 N° 40-62. Bogotá. D.C., Colombia

Adriana Vanegas Niño
Instituto de Errores Innatos del Metabolismo
Facultad de Ciencias
Pontificia Universidad Javeriana
Cra. 7 N° 40-62. Bogotá. D.C., Colombia

Patricia Landázuri
Laboratorio de Bioquímica y Genética
Facultad de las Ciencias de la Salud
Universidad del Quindío, Armenia, Colombia
Cra 15 Calle 12 Norte. Armenia, Quindío

Homero Sáenz
Instituto de Errores Innatos del Metabolismo
Facultad de Ciencias
Pontificia Universidad Javeriana
Cra. 7 N° 40-62. Bogotá. D.C., Colombia

Leonardo Lareo
Grupo de Investigación en Bioquímica Computacional
Departamento de Bioquímica. Facultad de Ciencias
Pontificia Universidad Javeriana
Cra. 7 N° 40-62. Bogotá. D.C., Colombia

Olga Yaneth Echeverri Peña
Instituto de Errores Innatos del Metabolismo
Facultad de Ciencias
Pontificia Universidad Javeriana
Cra. 7 N° 40-62. Bogotá. D.C., Colombia

Luis A. Barrera Avellaneda*
Instituto de Errores Innatos del Metabolismo
Facultad de Ciencias
Pontificia Universidad Javeriana
Cra. 7 N° 40-62. Bogotá. D.C., Colombia
E-mail: abarrera@javeriana.edu.co

* Corresponding author

Financial support: We acknowledge the financial support of Instituto Colombiano para la Ciencia y la Tecnología Francisco José de Caldas (COLCIENCIAS), Grant No. 1203-12-10410-192-2000 and Pontificia Universidad Javeriana, Bogotá, D.C., Colombia, Grant No. 120104-O-0101103.

Keywords: E. coli, glycation, human sulfatase, transient expression.

The recombinant human iduronate 2-sulfate sulfatase (hrIDS) was transiently and functionally active expressed in E. coli K12. The enzyme activity (crude extract) at 100ml and 400ml oscillated between 0.25 and 10.58nmol h^-1 mg^-1. The wide Western-blot peptide profile suggest that hrIDS is proteolitically processed "randomly" which agrees with the ultrafiltration assay in which the hrIDS activity was found in all fractions (< 30kDa, 30-100kDa and > 100kDa). No glycation sites were found by computer analysis of the hIDS sequence; discarding the possibility of marks for glycation and proteolytic processing.