Probing Cellular Processes for Productivity Enhancement in Hairy Root Cultures of C. roseus

从植物组织培养中生产药物和精细化学品（“次生代谢物”）多年来一直被认为是一个有价值的目标，但这项技术的潜力有限。一个主要的原因是细胞培养的遗传不稳定，以实现大规模的连续生产。一种新的植物培养体系是“毛状”根培养。PI已经在她的实验室建立了Catharanthus roseus（长春花）高产且稳定的“毛状”根培养。提出的研究包括调查毛状根培养物的代谢，以了解限制次生代谢物产生的因素。用高效液相色谱法（HPLC）对不同毛状根无性系在烧瓶培养中的生长动力学和生物碱产量进行监测。在灌注反应器中，采用31P和13C核磁共振（NMR）波谱法无创监测细胞培养物的生理状态和碳流。这种双重方法将增强对限制生物碱生产的代谢细胞过程的基本理解。

Production of pharmaceutical and fine chemicals ("secondary metabolites") from plant tissue culture has been recognized as a worthwhile objective for many years, but the potential of this technology has been limited. A major reason is the genetic instability of cell culture for continuous production on a large scale. A novel plant culture system is "hairy" root cultures. The PI has established in her laboratory productive and stable "hairy" root cultures for Catharanthus roseus (periwinkle). The proposed research involves investigating the metabolism of hairy root cultures in order to understand the factors which limit production of secondary metabolites. Growth kinetics and alkaloid production will be monitored by high performance liquid chromatography (HPLC) for different hairy root clones in flask cultures. The physiological state and carbon flows of the cell culture will be monitored noninvasively in a perfusion reactor by 31P and 13C Nuclear Magnetic Resonance (NMR) spectroscopy. This dual approach will enhance fundamental understanding of the metabolic cellular processes limiting alkaloid production.