P74
Improved production of the human recombinant enzime N-acetylgalactosamine-6-sulfatase in Escherichia coli
Monday, July 25, 2016
Grand Ballroom, 5th Fl (Sheraton New Orleans)
The enzyme replacement therapy is the main and sometimes only effective therapy for the treatment of some genetic diseases, as the lysosomal storage diseases. The Institute for the Study of Inborn Errors of Metabolism (IEIM) has been consolidated as a reference in Latin America for the diagnosis of such diseases, and it has demonstrated the production of active recombinant enzimes in different bacterial and yeast systems.
The enzyme N-acetylgalactosamine-6-sulfatase (GALNS) is an enzyme that catalyzes the degradation of the macromolecule keratan sulfate and chondroitin-6-sulfate. Mutations in GALNS cause a disease called Mucopolysaccharidosis 4A, an autosomal recessive lysosomal storage disease characterized by intracellular accumulation of these macromolecules. Key clinical features include short stature, skeletal dysplasia, dental anomalies, and corneal clouding. The IEIM has successfully produced GALNS in Escherichia coli; however the produced protein amounts were relatively low.
In this work, several strategies were investigated for the improvement and optimization of GALNS production. Approaches at the level of molecular and syntethic biology were successfully applied to increase GALNS production more than 12-fold. These approaches include improvement of gene transcription, and protein folding due to molecular chaperones and increases disulfide bond formation. All the strategies described in this work can be applied for the production of other human recombinant proteins.