Regulation of Cell Death for Increased Production of Acid α -glucosidase in Rice Cell Cultures

초록

Pompe disease caused by the deficiency of lysosomal acid α -glucosidase (GAA) enzyme is one of the lysosomal storage diseases. In this study, GAA enzyme was produced in transgenic rice cells using RAmy3D promoter which expresses target proteins by sugar depletion. However, when the rice cells are transferred to sugar-free medium for GAA production, this condition leads to autophagy due to nutrient depletion. In addition, cells go through apoptosis by various environmental stresses. These two types of cell death affect both the viability of cells and protein quality by proteolysis. In order to examine regulation effects of cell death in rice cell cultures for increased GAA productivity, four different chemicals (autophagy inhibitors: 3-methyladenine, concanamycin A/ autophagy enhancer: rapamycin/ apoptosis inhibitor: activated charcoal) were supplemented. Among them, addition of activated-charcoal exhibited the highest GAA productivity. Also, addition of concanamycin A increased GAA productivity while the other autophagy regulators did not show any effect. This result means that the regulation of cell death, especially inhibition of apoptosis, could enhance the production of target protein in rice cell cultures.