Abstract
Acid α-glucosidase (GAA) was produced in transgenic rice cell suspension cultures. In order to solve the problems using RAmy3D promoter for the efficient expression of GAA under sugar starvation condition, sorbitol was used as an alternative carbon source. Cell viability could be prolonged up to 27 days when sorbitol was present. Protein expression could also be enhanced. In addition, N-acetyl glucosamine, a hexokinase inhibitor, was used as a regulator for the RAmy3D promoter to shorten the incubation time with improved GAA expression. When sorbitol and N-acetyl glucosamine were used together, production of GAA was higher than that with sorbitol only. In order to optimize the combination condition, various experimental sets were designed and 60% increased protein could be obtained. In conclusion, optimized utilization of sorbitol as an alternative carbon source with N-acetyl glucosamine in transgenic rice cell suspension cultures was found to be the best condition for the enhanced expression of GAA using RAmy3D promoter.
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