Production and Characterization of Glucocerebrosidase in Transgenic Rice Cells

초록

Gaucher disease caused by deficiency of glucocerebrosidase (GCD) also known as β-glucosidase, is currently treated by enzyme replacement therapy.1) For GCD to be effective in this therapy, terminal mannose residues must be presented on the glycan chains to permit binding to the mannose receptors on macrophage which is target cell. In this study, GCD was expressed in transgenic rice cell suspension cultures and its characteristics were investigated. Plant-based systems were commonly used the production of glycoprotein, together with its targeting strategies generate GCD with terminal mannose structures for biological activity.2) The expression vector with rice RAmy3D promoter was constructed to express GCD and the production could be induced by sucrose starvation in medium.3) GCD cell lines were screened to confirm the fundamental characteristics of growth and production in callus form, respectively. After the induction, intracellular GCD extracted from the callus was characterized using Western blot. The calli expressing high levels of GCD was used to establish suspension cell lines. Growth properties of suspension cell lines were comparable to that of the other rice systems. The GCD was produced in suspension cultures after induction for 12~24 hours with sugar-free medium. The productivity of intracellular and extracellular GCD increased until 60~84 hours. Therefore, we can conclude that GCD could be successfully produced in transgenic rice suspension cells.