Neu5Ac衍生物和CMP-Neu5Ac衍生物高效合成技术的研究

项目名称： Neu5Ac衍生物和CMP-Neu5Ac衍生物高效合成技术的研究

项目编号： No.21202195

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 有机化学

项目作者： 吕迅

作者单位： 中国科学院微生物研究所

项目金额： 25万元

中文摘要： N-乙酰神经氨酸（Neu5Ac）、CMP-Neu5Ac及其衍生物具有重要生物学意义和药物研究价值，如何快速、大量地制备这些化合物是唾液酸研究领域中的一个关键基础问题。全细胞催化是目前最具工业化生产潜力的Neu5Ac合成方法，酶法合成则代表着CMP-Neu5Ac的主流制备技术，而国内外关于它们衍生物制备的研究却鲜有报道。本项目拟利用依托单位已建立的全细胞催化和酶法合成技术，同时结合有机合成，发展一种具有规模化生产潜力的Neu5Ac衍生物和CMP-Neu5Ac衍生物的高效合成方法，填补该领域中的空白。运用本项目的合成策略，对廉价易得的N-乙酰葡萄糖胺（GlcNAc）进行化学修饰后，再用两步生物催化反应可简便、快速地分别获得各种Neu5Ac衍生物和CMP-Neu5Ac衍生物。本项目将通过该方法大量制备（克级）结构多样的相关化合物，为后续的生物学和药学研究提供充足的物质储备。

中文关键词： N-乙酰神经氨酸；CMP-N-乙酰神经氨酸；全细胞催化；酶促合成；

英文摘要： Efficient and large-scale synthesis of N-acetyl neuraminic acid (Neu5Ac), CMP-Neu5Ac and their derivatives is very important because of the biological and pharmacological significance. The whole-cell catalysis is well studied and presents a new potential tool in industrial production of Neu5Ac. Convenient enzymatic synthesis of CMP-Neu5Ac is also recently well established by use of CMP-Neu5Ac synthase NeuA. However, the preparation of derivatives of Neu5Ac and CMP-Neu5Ac remains challenging due to their diverse and complex structures. Here we present an efficient synthetic strategy of the derivatives based on the combined use of the whole-cell catalysis and a chemoenzymatic approach. The proposed process contains (i) chemical modification of N-acetylglucosamine (GlcNAc) to produce a set of GlcNAc derivatives; (ii) whole-cell-catalyzed transformations of the Neu5Ac derivatives to Neu5Ac derivatives using a genetically engineered Escherichia coli strain we developed previously; and (iii) enzymatic preparation of the corresponding CMP-Neu5Ac derivatives from the GlcNAc derivatives by a recombinant NeuA overexpressed and purified from our another engineered E. coli strain. Our preliminary test have suggested that the whole-cell-catalyst as well as the recombinant NeuA exhibits a broad substrate tolerance, allowing s

英文关键词： N-acetyl neuraminic acid；CMP-N-acetyl neuraminic acid；whole-cell catalysis；chemoenzymatic synthesis；