Basic study on mass production of therapeutic proteins for serious disease by genetically engineered rice cell

利用基因工程水稻细胞大规模生产严重疾病治疗蛋白的基础研究

基本信息

批准号：
11650821
负责人：
TERASHIMA Masaaki
金额：
$ 2.3万
依托单位：
Osaka Prefecture University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

The following significant results were obtained from the research on production of therapeutic protein, α_1-antitrypsin (AAT) by genetically engineered rice cell.(1) Growth rate of genetically engineered cell was the same as that of non-transformed rice cell. Specific growth rate of rice cell, 0.3 d^<-1>, was of the same order of magnitude as those of other plant cells.(2) Oxygen transfer coefficient K_La was 7-8 h^<-1>, and aeration rate did not affect the growth rate of rice cell.(3) Ammonium was produced at cell growth phase and AAT production phase. However, ammonium did not affect the AAT productivity.(4) Adjustment of osmotic pressure between growth medium and production medium has favorable effective for AAT production.(5) AAT productivity was tripled using pyrvic acid as an altemative carbon source.(6) Continuous production was achieved in the continuous stirred tank reactor.(7) Cell density was increased up to 14 g-dry cell/L in the batch-type reactor.(8) Since maximum productivity of AAT at optimal condition was 25 mg/g-dry cell, at least 350 mg/L of AAT can be produced in batch-type reactor.The recombinant proteins can be rapidly induced by sugar depletion in the employed expression system. Our research strongly suggests that protein production system more efficient than microbial system can be constructed by the rice cell culture system. We hope the mass production system using plant cell culture can be achieved by exploring process-engineering research based on this fundamental research.

利用基因工程水稻细胞生产治疗性蛋白质α_1-抗胰蛋白酶（AAT）的研究取得了以下重要成果：（1）基因工程细胞的生长速度与非转化水稻细胞的生长速度相同。水稻细胞的速率为0.3 d^<-1>，与其他植物细胞处于同一数量级。 (2) 氧传递系数K_La为7-8 h^<-1>，通气量对水稻细胞的生长速度没有影响。(3)细胞生长期和AAT生产期均产生氨，但不影响AAT生产率。(4)生长培养基和生产培养基之间渗透压的调节。对AAT生产具有良好的效果。(5)使用丙酮酸作为替代碳源，AAT生产率提高了两倍。(6)在连续搅拌釜反应器中实现了连续生产。(7)细胞密度增加在间歇式反应器中，产量可达 14 g-干细胞/L。 (8) 由于最佳条件下 AAT 的最大生产率为 25 mg/g-干细胞，因此间歇式反应器中至少可以生产 350 mg/L 的 AAT我们的研究强烈表明，通过水稻细胞培养系统可以构建比微生物系统更有效的蛋白质生产系统。在此基础上探索工艺工程研究，可以实现植物细胞培养。