The attachment of polysaccharide chains to asparagine residues (N-linked glycosylation) confers unique properties to proteins, which are crucial for their solubility, stability, and enzymatic activity.

Escherichia coli, the most commonly used microorganism for heterologous protein expression, lacks glycosylation machinery. In contrast, Saccharomyces cerevisiae produces immunogenic glycans.

So, what if we give E. coli a tutorial and educate S. cerevisiae on how to produce human glycoproteins?

As proof of concept, we tried to produce the human Glucocerebrosidase (GCase), used in Gaucher’s disease treatment, in both organisms with the Man3GlcNAc2 glycosylation pattern.

Gaucher Disease (GD) is a genetic disease characterized by a deficiency of the enzyme beta-glycocerebrosidase, whose function is to degrade lipids called glycocerebrosides, leading to an impairment of lipid metabolism, which generates the accumulation of these lipids in cells such as macrophages. The main clinical manifestations of GD result from hematological, visceral, skeletal and neurological disorders, which originate from Gaucher cells, formed through the accumulation of glycocerebroside, and which are mostly found in organs such as the liver, spleen and bone marrow.

Currently, GD is divided into three types, which vary according to the clinical manifestations that individuals present.

Treatment for GD is mostly done through enzyme replacement therapy (ERT), and according to data from the Brazilian Ministry of Health, 96% of patients treated with ERT are given the enzyme glycocerebrosidase produced in laboratories and administered intravenously, while the remaining 4% use substrate synthesis inhibition (SSI). Therefore, the Glycosinside Out project aims to optimize the production of recombinant beta-glycocerebrosidase, leading to the easier production of this enzyme and, consequently, a reduction in costs for patients with GD.