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.