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.

从基因工程水稻细胞的热蛋白生产α_1-抗胰蛋白酶（AAT）的研究中获得了以下重要结果。（1）基因工程细胞的生长速率与未转化的水稻细胞的生长速率相同。水稻细胞的特定生长速率为0.3 d^<-1>，与其他植物细胞的生长量相同。（2）氧转移系数K_LA为7-8 H^<-1>，曝气速率不会影响水稻细胞的生长速率。（3）在细胞生长阶段和AAT生产阶段产生铵。然而，铵不会影响AAT的产生。（4）调整生长培养基和生产培养基之间的渗透压对AAT的产量有效。（5）使用Pyrvic Acid作为平均碳源将AAT产量增加了三倍。（6）在连续的打击储罐反应器中实现了连续的碳量。在最佳条件下为25 mg/g干细胞，可以在批处理型反应器中产生至少350 mg/L的AAT。重组蛋白可以通过糖耗尽在员工表达系统中迅速诱导。我们的研究强烈表明，蛋白质生产系统比微生物系统更有效，可以由水稻细胞培养系统构建。我们希望可以通过基于这项基本研究来探索工艺工程研究来实现使用植物细胞培养的大众生产系统。