Laboratory for Biomaterials, Institute for Biotechnology and Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany.
Laboratory for Biomaterials, Institute for Biotechnology and Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany.
J Biotechnol. 2017 Sep 20;258:51-55. doi: 10.1016/j.jbiotec.2017.03.025. Epub 2017 Mar 24.
The broad substrate spectrum of UDP-sugar pyrophosphorylases from plant salvage pathways is of high interest for the synthesis of expensive nucleotide sugars by straightforward enzyme cascade reactions in combination with monosaccharide kinases. We here present a new UDP-sugar pyrophosphorylase from Hordeum vulgare with favorable biochemical properties like broad pH and temperature tolerances as well as a broad substrate spectrum and high synthesis stability. Enzyme properties were determined and reaction conditions were optimized by high-through-put multiplexed capillary electrophoresis analysis. In combination with a galactokinase UDP-α-d-galactose (UDP-Gal) was efficiently synthesized with a space-time-yield of 17g/L*h for full conversion of 10mM substrate within 20min by 1.2U of each enzyme.
植物补救途径中的 UDP-糖焦磷酸化酶具有广泛的底物谱，通过与单糖激酶结合的直接酶级联反应来合成昂贵的核苷酸糖，这引起了人们的极大兴趣。在这里，我们介绍了一种来自大麦的新型 UDP-糖焦磷酸化酶，它具有良好的生化特性，如宽 pH 和温度耐受性以及广泛的底物谱和高合成稳定性。通过高通量多重毛细管电泳分析来确定酶的性质并优化反应条件。与半乳糖激酶 UDP-α-d-半乳糖 (UDP-Gal) 结合使用，在 20 分钟内可有效合成 UDP-α-d-半乳糖，时空产率为 17g/L*h，10mM 底物完全转化，每种酶用量为 1.2U。
