Developing chemoenzymatic strategies, enzymes, and kits for accessible and affordable gangliosides

项目摘要 神经节苷脂类似于细胞中的生物触角，在多种生物学行为中发挥关键作用。 过程（即病原体感染、细胞间通讯、炎症）。神经保护和神经恢复 已经探索了神经节苷脂在帕金森病和其他疾病中的治疗应用， 神经退行性疾病尽管神经节苷脂具有如此重要的作用，但神经节苷脂的可及性有限。 目前大多数市售的神经节苷脂是从动物脑中获得的，这可能是一种新的方法。 关注未来的治疗应用。神经节苷脂的这种有限的可用性进一步受到以下因素的困扰： 基于不同的糖和脂肪酸的多样性。这里介绍的技术涵盖了 近十年的研究，其中复杂的神经节苷脂是在一个单一的反应容器中合成， 一种简单的纯化技术。这种生物合成工具打开了急需的神经节苷脂的可用性， 研究并支持其治疗用途。在这份快速通道提案中，我们将应对以下方面的一些重大挑战： 将这种原型一锅多酶（OPME）技术转变为大规模商业化。第一 挑战是制造起始材料（糖基转移酶、补充酶和 乳糖基鞘氨醇）以降低生产神经节苷脂的成本。现有的其他改进 糖基转移酶将通过提高催化活性来进一步降低生产神经节苷脂的成本负担。 酶的效率。最后，将探索OPME Scalpers用于制造各种 神经节苷脂一项研究将导致几种商业产品，从纯化的酶，微型化， 糖基化试剂盒和几种具有高商业价值的关键神经节苷脂。据报道，GM1神经节苷脂 延迟帕金森病的发作，临床试验可能需要无动物神经节苷脂。一种小型化 将提供试剂盒作为研究人员合成自己定制的神经节苷脂的工具。的 糖基转移酶和OPME试剂盒允许广泛的科学界获得这些简单的合成 其他寡糖、糖缀合物和糖蛋白的工具。通过扩大OPME并降低 通过这些产品的成本，将获得关于神经节苷脂生物学功能的额外信息， 加快神经节苷脂为基础的诊断和治疗的发展。一个积极的反馈回路，其中 较低的成本将增加采购量，这将为更多的研究提供资金，并将导致 糖基转移酶、OPME试剂盒和高纯度、无动物源性神经节苷脂的市场。

Project Summary Gangliosides, analogous to biological antennae in cells, are involved in critical roles across multiple biological processes (i.e. pathogen infection, cell-cell communication, inflammation). Neuroprotective and neurorestorative functions of gangliosides have been explored for their therapeutic application in Parkinson’s disease and other neurodegenerative diseases. Despite such important role of gangliosides, there is limited access of gangliosides. Majority of the current commercially available gangliosides are obtained from animal brains, which is likely a concern for future therapeutic applications. Such limited availability of gangliosides is further beset by the diversity based on different sugars and fatty acids. The technologies presented here cover an amalgamation of nearly a decade of research, where complex ganglioside is synthesized in a single reaction vessel followed by a simple purification technique. This biosynthetic tool opens the availability of critically needed gangliosides in research and support its therapeutic uses. In this fast track proposal, we will address some major challenges in transitioning this prototypic one-pot multienzyme (OPME) technology for large scale commercialization. First challenge is manufacturing of starting materials (glycosyltransferases, supplemental enzymes and lactosylsphingosine) to reduce the cost to manufacture gangliosides. Additional improvements of existing glycosyltransferases will further reduce the cost burden to manufacture gangliosides by improving catalytic efficiencies of the enzymes. Lastly, OPME scaleability will be explored for manufacturing the various gangliosides. A study will result in several commercial products, ranging from purified enzymes, miniaturized glycosylation kits and several key gangliosides with high commercial value. GM1 ganglioside has been reported to delay onset of Parkinson disease, and clinical trials will likely require animal-free gangliosides. A miniaturized kit will be provided as a tool for researchers to synthesize their own customized gangliosides. The glycosyltransferases and the OPME kits allow the broad scientific community to access these simple synthetic tools for other oligosaccharides, glycoconjugates, and glycoproteins. By expanding this OPME and lowering the cost of these products, additional information on the biological functions of gangliosides will be gained which will accelerate the development of ganglioside-based diagnostics and therapeutics. A positive feedback loop, where lower cost will increase purchase volume, which will feed additional studies and will lead to the further growth of the market for the glycosyltransferases, OPME kits, and highly pure, animal-free gangliosides.