Harvesting specific plant metabolites from hairy root cultures using magnetized n

使用磁化n从毛状根培养物中收获特定的植物代谢物

• 批准号: 8712853
• 负责人: JOHN M. LITTLETON
• 金额: $ 18.98万
• 依托单位: NAPROGENIX, INC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8712853
• 关键词: Address; Area; Binding; Biological Factors; Biotechnology; Cell Culture Techniques; Cell Survival; Cells; Characteristics; Chemical Engineering; Chemicals; Collaborations; Complex Mixtures; Devices; Drug Industry; Endocytosis; Engineering; Exposure to; Faculty; Fermentation; Flavonoids; Funding; Genetically Modified Plants; Harvest; High Pressure Liquid Chromatography; Individual; Industry; Kentucky; Licensing; Life; Mammalian Cell; Marketing; Measures; Medicine; Methods; Nutrient; One-Step dentin bonding system; Pharmacologic Substance; Phase; Plant Roots; Plants; Preparation; Process; Production; Protocols documentation; Quercetin; Research; Research Personnel; Schedule; Silicon Dioxide; Small Business Technology Transfer Research; Solidago; Specificity; Surface; System; Taxoids; Technology; Testing; Time; Tissues; Universities; Vinca Alkaloids; chemotherapeutic agent; commercialization; cytotoxicity; design; extracellular; improved; in vitro testing; magnetic field; microbial; mutant; nanoparticle; novel; prototype; public health relevance; radioligand; response; tissue culture

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.

产品说明：植物产生多种有价值的生物活性代谢物，但这些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且昂贵。申请人公司Naprogix Inc开发了一种技术，用于增加突变植物细胞培养物中特定生物活性代谢物的产量，有望减轻这一问题。然而，这种方法或使用遗传修饰的植物细胞的任何其他方法的价值受到为了提取所需产物而浪费性破坏植物细胞的必要性的限制。 如果可以从连续培养的细胞中收获产物，则该过程将更有效。纳米颗粒通过内吞作用被细胞摄取，随后被胞吐。它们也可以被设计成吸附特定的化学物质。这表明，特别工程化（功能化和磁化）的纳米粒子可用于反复“收获”来自活植物细胞培养物的特定代谢物。该公司的主要项目之一是在一枝黄花毛状根培养中产生新型黄酮类化合物。申请人已经表明，官能化的二氧化硅纳米颗粒吸附类似的类黄酮，并且它们被吸收并从这些植物细胞培养物中除去，而不损害培养物的活力。现在的目标是设计最佳功能化的磁化二氧化硅纳米颗粒（由肯塔基州大学的合作者），从这些毛状根中收获特定的黄酮类化合物。 阶段I的具体目的是显示（a）暴露于这些纳米颗粒将类黄酮从毛状根的细胞内区室转移到细胞外介质（B）这些细胞外磁化的纳米颗粒可以在磁场中浓缩（c）在这些纳米颗粒通过植物细胞后，可以从这些纳米颗粒洗脱和收集大量的类黄酮。如果这是成功的，那么在第二阶段，申请人将设计用于从优化的植物细胞培养物中收获其他生物活性代谢物的测试纳米颗粒。这些将包括非常高价值的化学治疗剂，如紫杉烷和长春花生物碱。如果成功的话，这种方法将对利用植物细胞培养生产和分离高价值的天然产品产生重大影响。纳米颗粒和应用于植物细胞的收获技术是可申请专利的，对申请公司和大学具有重大的商业意义。

项目成果

Mechanism of Mesoporous Silica Nanoparticle Interaction with Hairy Root Cultures during Nanoharvesting of Biomolecules.

生物分子纳米收获过程中介孔二氧化硅纳米粒子与毛状根培养物相互作用的机制。

• DOI: 10.1002/adbi.202000173
• 发表时间: 2021
• 期刊: Advanced biology
• 影响因子: 3.7
• 作者: Khan,MdArif;Fugate,Madeleine;Rogers,DennisT;Sambi,Jatinder;Littleton,JohnM;Rankin,StephenE;Knutson,BarbaraL
• 通讯作者: Knutson,BarbaraL

Strategy for Conjugating Oligopeptides to Mesoporous Silica Nanoparticles Using Diazirine-Based Heterobifunctional Linkers.

• DOI: 10.3390/nano12040608
• 发表时间: 2022-02-11
• 期刊: Nanomaterials (Basel, Switzerland)
• 作者: Khan MA;Ghanim RW;Kiser MR;Moradipour M;Rogers DT;Littleton JM;Bradley LH;Lynn BC;Rankin SE;Knutson BL
• 通讯作者: Knutson BL