Construction of Processes for Production and Separation of Pigments Originated from Plant

自古以来，植物就在为人类提供多种生理活性物质方面发挥着重要作用。此外，植物一直是工业规模生产饮料、药品、颜料和食品添加剂的重要原料。植物细胞衍生产品的成功商业应用需要了解植物细胞遗传学、生理学、代谢调节和生物过程工程的原理。在利用植物毛状根生产植物色素的培养体系中，仍有许多问题需要解决。本项目致力于构建适合植物毛状根生产植物色素的培养体系。为解决毛状根培养中与工程相关的一些关键问题，开发生产工艺，对红甜菜和茜草毛状根进行了研究。为了改善培养条件，提出了提高目标代谢物产量的策略。…此外，在红甜菜毛状根的培养中，研究了基质改良对色素产量的影响，结果表明，磷酸盐是促进色素形成的重要成分。在茜草毛状根的培养中，碳源和氮源是重要的影响因素，在以果糖为碳源的无铵培养中，色素产量显著提高。此外，在毛状根生长的动力学方面，基于根尖生长点的线性延伸和侧向分枝的模型使得模拟和表征不同毛状根的生长成为可能。为了发展红甜菜和茜草毛状根高效生产色素的培养操作，进行了与生长和色素释放操作相关的重复培养。从茜草毛状根中提取的色素被定义为茜素、芦荟素及其苷类。为了获得目标产物茜素的较高得率，采用了有机溶剂萃取法进行酶促反应。采用β-古尔糖苷酶对茜素糖苷进行选择性水解，并选择正己烷作为茜素的提取方法。较少

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. … More 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. Less