Phosphomannose isomerase/GDP-mannose pyrophosphorylase from Pyrococcus furiosus: a thermostable biocatalyst for the synthesis of guanidinediphosphate-activated and mannose-containing sugar nucleotides
Phosphomannose isomerase/GDP-mannose pyrophosphorylase from Pyrococcus furiosus: a thermostable biocatalyst for the synthesis of guanidinediphosphate-activated and mannose-containing sugar nucleotides
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DOI：
10.1039/b822794b
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发表时间：
2009-01-01
影响因子：
3.2
通讯作者：
Pohl, Nicola L. B.
中科院分区：
文献类型：
--
作者：
Mizanur, Rahman M.;Pohl, Nicola L. B.
本文分析了从激烈火球菌（Pyrococcus furiosus）DSM 3638中获得的重组双功能磷酸甘露糖异构酶/GDP-甘露糖焦磷酸化酶（manC）的化学功能及其在合成胍二磷酸己糖和一系列核苷酸二磷酸甘露糖中的应用。这种酶在其核苷酸三磷酸（NTP）和糖-1-磷酸接受方面异常混杂。它接受所有五种天然存在的NTP（ATP，CTP，GTP，dTTP和UTP）和一系列糖-1-磷酸（葡萄糖-，甘露糖-，半乳糖-，葡糖胺-，N-乙酰葡糖胺-和岩藻糖-1-磷酸）。一个截短的GDP-甘露糖焦磷酸化酶结构域的整个长度的酶表现出几乎100倍少的糖核苷酸转移酶活性，只有GTP和甘露糖1-磷酸作为底物。这种古细菌酶的温度稳定性和固有的广泛底物耐受性使其成为快速化学酶促合成一系列天然和非天然糖核苷酸的有效试剂，这些天然和非天然糖核苷酸仅通过化学方法制备具有挑战性。
Herein we present an analysis of the chemical function of a recombinant bifunctional phosphomannose isomerase/GDP-mannose pyrophosphorylase (manC) from Pyrococcus furiosus DSM 3638 and its use in the synthesis of guanidinediphospho-hexoses and a range of nucleotidediphospho-mannoses. This enzyme is unusually promiscuous in both its nucleotide triphosphate (NTP) and sugar-1-phosphate acceptance. It accepts all five naturally occurring NTPs (ATP, CTP, GTP, dTTP and UTP) and a range of sugar-1-phosphates (glucose-, mannose-, galactose-, glucosamine-, N-acetylglucosamine- and fucose-1-phosphate). A truncated GDP-mannose pyrophosphorylase domain of the whole length enzyme showed almost 100-fold less sugar nucleotidyltransferase activity with only GTP and mannose 1-phosphate as substrates. The temperature stability and inherently broad substrate tolerance of this archaeal enzyme make it an effective reagent for the rapid chemoenzymatic synthesis of a range of natural and unnatural sugar nucleotides that are challenging to make by chemical means alone.