Yarema K J
Whitaker Biomedical Engineering Institute, Whiting School of Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
Biotechniques. 2001 Aug;31(2):384-93. doi: 10.2144/01312rv02.
This review discusses new directions in the emerging field of carbohydrate engineering. Specifically, it describes substrate-based methodologies that are complementary to the recombinant DNA techniques that now dominate metabolic and cellular engineering endeavors. A substrate-based approach consists of intercepting a biosynthetic pathway with an unnatural analog of a metabolic intermediate. The unnatural compound competes with the endogenous substrate for biosynthetic incorporation into a cellular component by action of the natural enzymes of the cell. The incorporation of the unnatural compound into the cellular architecture can directly modulate cellular properties and biological processes. Alternatively, a molecular handle can be included in the design of the unnatural substrate that allows further elaboration upon reaction with an externally delivered reagent. The sialic acid biosynthetic pathway is presented as a model system to illustrate both the practical aspects and theoretical considerations of a substrate-based cellular engineering approach. Specific applications of carbohydrate-based cell surface engineering include chemical construction of new glycosylation patterns on cells, new approaches to targeting tumor cell with either diagnostic or therapeutic agents, and installation of novel receptors on cells for facilitating viral-mediated gene delivery.
本综述讨论了碳水化合物工程这一新兴领域的新方向。具体而言，它描述了基于底物的方法，这些方法是对目前主导代谢和细胞工程研究的重组DNA技术的补充。基于底物的方法包括用代谢中间体的非天然类似物拦截生物合成途径。这种非天然化合物通过细胞天然酶的作用与内源性底物竞争，以生物合成方式掺入细胞成分中。将非天然化合物掺入细胞结构可直接调节细胞特性和生物过程。或者，可以在非天然底物的设计中加入一个分子手柄，以便在与外部递送的试剂反应后进行进一步修饰。唾液酸生物合成途径作为一个模型系统，用于说明基于底物的细胞工程方法的实际方面和理论考量。基于碳水化合物的细胞表面工程的具体应用包括在细胞上化学构建新的糖基化模式、用诊断或治疗剂靶向肿瘤细胞的新方法，以及在细胞上安装新型受体以促进病毒介导的基因递送。
