2000 Fiscal Year Final Research Report Summary
Basic study on mass production of therapeutic proteins for serious disease by genetically engineered rice cell
| Project/Area Number |
11650821
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | Osaka Prefecture University |
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Principal Investigator |
TERASHIMA Masaaki Osaka Prefecture Univ., Chem.Eng., Associate Prof., 工学研究科, 助教授 (30172092)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIDA Hiroyuki Osaka Prefecture Univ., Chem.Eng., Professor, 大学院・工学研究科, 教授 (50081360)
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| Project Period (FY) |
1999 – 2000
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| Keywords | rice cell / recombinant protein / antitrypsin / high density culture / specific growth rate / continuous culture / secretion / therapeutic protein |
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| Research Abstract |
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.
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