KINO-OKA Masahiro OSAKA UNIV., DEPT.OF CHEM.SCI.ENG., RESEARCH ASSOCIATE, 大学院・基礎工学研究科, 助手 (40234314)
MASAWAKI Teruyuki OSAKA UNIV., DEPT.OF CHEM.SCI.ENG., ASSOCIATE PROFESSOR, 大学院・基礎工学研究科, 助教授 (10209421)
TAYA Masahito OSAKA UNIV., DEPT.OF CHEM.SCI.ENG., PROFESSOR, 大学院・基礎工学研究科, 教授 (60144127)
|Budget Amount *help
¥7,800,000 (Direct Cost : ¥7,800,000)
Fiscal Year 1997 : ¥3,100,000 (Direct Cost : ¥3,100,000)
Fiscal Year 1996 : ¥4,700,000 (Direct Cost : ¥4,700,000)
Plants have playd an important role in supplying human being with many physiologically active substances since ancient times. Moreover, plants have been important as a raw materials for beverages, medicines, pigments and food additives on an industrial scale. The success in commercial use of plant cell-derived products requires knowledge of the principles of plant cell genetics, physiology, metabolic regulation, and bioprocess engineering. Many problems remain to be solved in culture systems which facilitate high yields of both biomass and the desired metabolites.
This project dealt with construction of culture systems suitable to the production of plant-derived pigments by using plant hairy roots. To resolve some key problems related with engineering aspects in hairy root cultures and to develop production process, red beet and madder hairy roots were examined.
1. For the improvement of culture conditions, the strategy for enhancement of productivity of desired metabolite was proposed.
The effect of medium modification on pigment productivity was investigated in the culture of red beet hairy roots and phosphate was identified as an important constituent which stimulated pigment formation. In the culture of madder hairy roots, carbon and nitrogen sources were important factors and the productivity of pigment was drastically enhanced in the culture using ammonium-free medium containing fructose as a carbon source. Moreover, concerning the kinetics of hairy root proliferation, the model based upon the linear extension and lateral branching of the growing points at root tip made it possible to simulate and characterize the growth of various hairy roots.
2. For the development of the culture operation for the effective production of pigment by red beet and madder hairy roots, repeated cultures associated with operations of growth and pigment release were performed. The pigments from madder hairy roots were defined as the alizarin, lucidin and their glycosides. To achieve the higher yield for the alizarin which is target product, the enzymatic reaction accompanied with extraction by organic solvent was performed. beta-gulcosidase was suitable to selective hydrolysis of alizarin glycoside and hexane was chosen for the extraction of alizarin.