1999 Fiscal Year Final Research Report Summary
Development of useful metabolites production system by protoplasts with artificial cell wall.
| Project/Area Number |
09555248
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
生物・生体工学
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| Research Institution | University of Tsukuba |
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Principal Investigator |
TANAKA Hideo Institute of Applied Biochemistry, University of Tsukuba Professor, 応用生物化学系, 教授 (40015657)
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| Co-Investigator(Kenkyū-buntansha) |
AOYAGI Hideki Institute of Applied Biochemistry, University of Tsukuba Assistant Professor, 応用生物化学系, 講師 (00251025)
OGBONNA James Institute of Applied Biochemistry, University of Tsukuba Associate Professor, 応用生物化学系, 助教授 (00261777)
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| Project Period (FY) |
1997 – 1999
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| Keywords | artificial cell wall / plant protoplast / microbial protoplast / useful metabolites production / bioreactor / elicitor / alginate / inhibitor of cell wall synthesis |
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| Research Abstract |
Study on development of useful metabolites production system by microbial and plant protoplasts with artificial cell wall were carried out. The following results were obtained.
(1) Discovery that alginate acts as an elicitor and can be used as an artificial cell wall.
Various studies on the functions of alginate showed that alginate acts as an elicitor in promoting the production of many useful compounds by plant cells. The basic analysis of action mechanism of alginate was carried out, and it was postulated that there was a recognition site for alginate on cell membrane. It was found that in the protoplast which has no cell wall, the sensitivity of the alginate was high, and that the elicitor-effect remarkably increased. The elicitor-effect of alginate increased significantly when HィイD22ィエD2OィイD22ィエD2 and CaィイD12+ィエD1 were added simultaneously.
(2) Development of simple method for preparation of microbial and plant protoplasts with artificial cell wall.
After immobilization of cells in al
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ginate, the beads containing the immobilized cells were treated with cell wall digesting enzyme and protoplasts were formed inside the beads. This is a very simple and rapid method for producing both microbial and plant protoplasts with artificial cell walls.
(3) The development of useful compound production system by the plant protoplasts with artificial cell wall.
Catharanthus roseus protoplast were immobilized in alginate gel with the elicitor function as an artificial cell wall, and used for indole alkaloids production. In comparison with normal cells, the indole alkaloids production increased more than 30 times. Also system for cultivation of Wasabia japonica protoplast with artificial cell wall and with addition of HィイD22ィエD2OィイD22ィエD2 was developed. In this study, HィイD22ィエD2OィイD22ィエD2 and alginate acted synergistically and chitinase production increased by more than one order when comparison with normal cells.
(4) Development of useful compound production system by the microbial protoplast with artificial cell wall.
Protoplasts were prepared from yeast cells for production of invertase. Alginate gel with high mass transfer capacity was used as the artificial cell wall. When the protoplasts were cultivated in bubble column reactor, the invertase production increased much larger than that of the cells. Less
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