Application of Anoxic Conditions with Various Carbon Sources in Glycoengineered Rice Cell Cultures

초록

For the production of biopharmaceutical proteins, plant expression systems provide several advantages such as post-translational modifications and safety [1]. To achieve the high titer of target proteins, RAmy3D promoter has been used in plant cell cultures. Since RAmy3D promoter is induced by sugar starvation, cell death is inevitable with negative effects on product quality and productivity [2]. For delaying cell death, anoxic conditions with the addition of various carbon sources can be applied in plant cell cultures [3]. In this study, glycoengineered rice cells were cultured in anoxic conditions. Three different carbon sources (sucrose, glucose, fructose) were added into culture medium. In anoxic conditions, consumption rates of sugars were slower than those of control cultures. Especially, at day 6, improvement of cell viability was observed by the addition of sucrose, glucose and fructose in anoxic conditions (78, 64.2 and 53.1%, respectively) compared to control (32.3, 24.8 and 22.6%, respectively) and sugar-free conditions (8.2%). In addition, it was found that protease activity was decreased in anoxic conditions. In conclusion, anoxic conditions with the addition of carbon sources could be applied in glycoengineered rice cell cultures for improving cell viability with the maintenance of product quality.