Utilization of Kifunensine and Sorbitol for Enhanced Production of Glyco-engineered Acid α-Glucosidase in Tansgenic Rice Cell Cultures

초록

Pompe disease is caused by the deficiency of acid α-glucosidase (GAA) and can be treated with enzyme replacement therapy (ERT) which allows targeting of lysosomal enzymes to the lysosome by mannose-6-phosphate (M6P) receptor. Kifunensine as a glycosylation inhibitor can be added to culture medium to obtain high-mannose type glycan for targeting M6P. However, it negatively affects protein production because of ER stress. Sugar alcohols as alternative carbon sources may protect from both oxidative stress and reduction of protein production. In this study, kifunensine was added to culture medium for glycolengineering in transgenic rice cells, which makes possible to obtain high-mannose glycan. When kifunensine was added, GAA production was decreased 1.18-fold compared to that of control condition. However, simultaneous addition of sorbitol with kifunensine increased the GAA production 1.4-fold. Also, addition of kifunensine was found to be effective to obtain high-mannose glycan. In conclusion, kifunensine is known to be a glycosylation inhibitor and sorbitol can be used as an alternative carbon source for enhancing GAA production in transgenic plant cell culture.