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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 13, No. 3, 2010
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Bioline Code: ej10027
Full paper language: English
Document type: Research Article
Document available free of charge
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Electronic Journal of Biotechnology, Vol. 13, No. 3, 2010
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Human sulfatase transiently and functionally active expressed in E. coli K12
Poutou-Piñales, Raúl A.; Niño, Adriana Vanegas; Landázuri, Patricia; Sáenz, Homero; Lareo, Leonardo; Peña, Olga Yaneth Echeverri & Avellaneda, Luis A. Barrera
Abstract
The recombinant human iduronate 2-sulfate sulfatase (hrIDS) was transiently and functionally active expressed in E. coli K12. The enzyme activity (crude extract) at 100 ml and 400 ml oscillated between 0.25 and 10.58 nmol 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.
Keywords
E. coli, glycation, human sulfatase, transient expression.
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