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형질전환 벼 세포 배양에서 Sorbitol과 Hexokinase 저해제 조합 전략을 통한 Acid α-glucosidase 생산성 증대
정창현(Chang-Hyun Jung),최홍열(Hong-Yeol Choi),복다은(Da Eun Bok),양유정(Yu Jeong Yang),이현아(Hyeon Ah Lee),이지아(Ji Ah Lee),최은수(Eun Soo Choi),김동일(Dong-Il Kim) 한국생물공학회 2021 KSBB Journal Vol.36 No.2
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.