Harvesting specific plant metabolites from hairy root cultures using magnetized n

植物产生多种有价值的生物活性代谢物，但这些代谢物通常以低浓度存在于野生型植物中。这使得这些化合物的分离和纯化变得复杂和昂贵。申请公司Naprogenix Inc .已经开发出一种技术，可以提高突变植物细胞培养中特定生物活性代谢物的产量，有望减轻这一问题。然而，这种方法的价值，或者任何其他使用转基因植物细胞的方法，都受到为了提取所需产品而对植物细胞进行浪费破坏的必要性的限制。

DESCRIPTION: Plants produce a wide variety of valuable bioactive metabolites, but these are commonly present in low concentrations in the wild-type plant. This makes the separation and purification of these compounds complicated and expensive. The applicant company, Naprogenix Inc, has developed a technology for increasing the yields of specific bioactive metabolites in mutant plant cell cultures, which promises to lessen this problem. However, the value of this, or any other approach using genetically-modified plant cells, is limited by the necessity of the wasteful destruction of the plant cells in order to extract the required products. The process would be more efficient if products could be harvested from cells in continuous culture. Nanoparticles are taken up by cells by endocytosis, and subsequently exocytosed. They can also be engineered to adsorb specific chemicals. This suggests that specifically-engineered (functionalized and magnetized) nanoparticles could be used to repeatedly "harvest" specific metabolites from living plant cell cultures. One of the company's major projects is designed to generate novel flavonoids in goldenrod hairy root cultures. The applicants have shown that functionalized silica nanoparticles adsorb similar flavonoids and that they are taken up and removed from these plant cell cultures without compromising the viability of the cultures. The objective is now to engineer optimally-functionalized magnetized silica nanoparticles (by collaborators at the University of Kentucky) to harvest specific flavonoids from these hairy roots. The specific aims of phase I are to show that (a) exposure to these nanoparticles translocate flavonoids from the intracellular compartment of hairy roots to the extracellular medium (b) these extracellular magnetized nanoparticles can be concentrated in a magnetic field (c) significant amounts of flavonoids can be eluted and collected from these nanoparticles following their passage through plant cells. If this is successful, then, in phase II, the applicants will design ad test nanoparticles for harvesting other bioactive metabolites from optimized plant cell cultures. These will include very high value chemotherapeutic agents, such as the taxoids and vinca alkaloids. If successful this approach should make a major impact on the use of plant cell cultures for the production and isolation of high value natural products. The nanoparticles, and the harvesting technologies as applied to plant cells, are patentable, and have major commercial implications for the applicant company and the University.