Comparing substrate specificity of two UDP-sugar pyrophosphorylases and efficient one-pot enzymatic synthesis of UDP-GlcA and UDP-GalA.

Comparing substrate specificity of two UDP-sugar pyrophosphorylases and efficient one-pot enzymatic synthesis of UDP-GlcA and UDP-GalA.
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比较两个UDP糖焦磷酸酶的底物特异性以及UDP-GLCA和UDP-GALA的有效的一锅酶促合成。

DOI:
10.1016/j.carres.2015.04.001
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发表时间:
2015-06-26
影响因子:
3.1
通讯作者:
Li, Lei
Li, Lei
中科院分区:
化学3区
文献类型:
--
作者:
Guo, Yuxi;Fang, Junqiang;Li, Tiehai;Li, Xu;Ma, Cheng;Wang, Xuan;Wang, Peng G.;Li, Lei

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尿苷-5‘-二磷酸-葡萄糖醛酸(UDP-GLCA)和UDP-半乳糖醛酸(UDP-GALA)是唯一的羧酸形成的糖核苷酸,是参与多种细胞成分生物合成的关键前体。这些成分的可获得性有限,阻碍了开发简便地合成糖胺多糖等生物活性多糖的有效方法。在目前的研究中,我们从拟南芥和婴儿双歧杆菌ATCC15697(BiUSP)中对两种UDP-糖焦磷酸化酶(AtUSP)进行了生化表征,并比较了它们对一组糖-1-磷酸及其衍生物的活性。这两种酶对Glca-1-磷酸表现出显著的焦磷酸化活性,而AtUSP对Gala-1-磷酸也表现出类似的活性。通过结合单糖-1-磷酸酶,我们开发了一种高效、简便的一锅三酶法,可以快速获得数百毫克的UDP-GLCA和UDP-GALA。
Uridine 5’-diphosphate-glucuronic acid (UDP-GlcA) and UDP-galacturonic acid (UDP-GalA), the unique carboxylic acid-formed sugar nucleotides, are key precursors involved in the biosynthesis of numerous cell components. Limited availability of those components has been hindering the development of efficient ways towards facile synthesis of bioactive glycans such as glycosaminoglycans. In current study, we biochemically characterized two UDP-sugar pyrophosphorylases from Arabidopsis thaliana (AtUSP) and Bifidobacterium infantis ATCC15697 (BiUSP), and compared their activities towards a panel of sugar-1-phosphates and derivatives. Both enzymes showed significant pyrophosphorylation activities towards GlcA-1-phosphate, and AtUSP also exhibited comparable activity towards GalA-1-phosphate. By combining with monosaccharide-1-phosphate kinases, we have developed an efficient and facile one-pot three-enzyme approach to quickly obtain hundreds milligrams of UDP-GlcA and UDP-GalA.
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