Kuberan Balagurunathan, Lech Miroslaw Z, Beeler David L, Wu Zhengliang L, Rosenberg Robert D
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Nat Biotechnol. 2003 Nov;21(11):1343-6. doi: 10.1038/nbt885. Epub 2003 Oct 5.
Heparan sulfate (HS) proteoglycans are crucial to numerous biological processes and pathological conditions, but to date only a few HS structures have been synthesized and characterized with regard to structure-function relationships. Because HS proteoglycans are highly diverse in structure, there are substantial limitations on their synthesis by classical chemical means, and thus new methods to rapidly assemble bioactive HS structures are needed. Here we report the biosynthesis of bioactive HS oligosaccharides using an engineered set of cloned enzymes that mimics the Golgi apparatus in vitro. We rapidly and efficiently assembled the antithrombin III-binding pentasaccharide in just 6 steps, in contrast to the approximately 60 steps needed for its chemical synthesis, with an overall yield at least twofold greater and a completion time at least 100 times faster than for the chemical process.
硫酸乙酰肝素（HS）蛋白聚糖对众多生物学过程和病理状况至关重要，但迄今为止，关于结构-功能关系，仅合成并表征了少数几种HS结构。由于HS蛋白聚糖结构高度多样，用传统化学方法合成它们存在很大局限性，因此需要新方法来快速组装具有生物活性的HS结构。在此，我们报告了利用一组经工程改造的克隆酶在体外模拟高尔基体来生物合成具有生物活性的HS寡糖。我们仅用6步就快速高效地组装出了抗凝血酶III结合五糖，相比之下其化学合成大约需要60步，总体产率至少高出两倍，完成时间比化学过程至少快100倍。
