6815-91-4Relevant articles and documents
Gram-scale production of sugar nucleotides and their derivatives
Li, Shuang,Wang, Shuaishuai,Wang, Yaqian,Qu, Jingyao,Liu, Xian-Wei,Wang, Peng George,Fang, Junqiang
supporting information, p. 2628 - 2633 (2021/04/21)
Here, we report a practical sugar nucleotide production strategy that combined a high-concentrated multi-enzyme catalyzed reaction and a robust chromatography-free selective precipitation purification process. Twelve sugar nucleotides were synthesized on a gram scale with a purity up to 98%.
A Kinase-Independent One-Pot Multienzyme Cascade for an Expedient Synthesis of Guanosine 5′-Diphospho-d-mannose
Pfeiffer, Martin,Bulfon, Dominik,Weber, Hansjoerg,Nidetzky, Bernd
, p. 3809 - 3816 (2016/12/16)
Biomimetic synthesis routes towards the important natural d-mannosyl donor guanosine 5′-diphospho-d-mannose (GDP-Man) rely on kinase-catalyzed nucleotide triphosphate (NTP)-dependent phosphorylations of d-mannose (Man), to give d-mannose 6-phosphate or α-d-mannose 1-phosphate (αMan 1-P) as an intermediate product. A GDP-Man synthesis not requiring the kinase/NTP system would be practical and cost-effective. Here, we have developed a multienzyme cascade towards GDP-Man, characterized in that αMan 1-P was obtained by a diastereoselective phosphatase-catalyzed phosphorylation of Man. α-d-Glucose 1-phosphate (αGlc 1-P), prepared in situ through phosphorylase-catalyzed conversion of sucrose in the presence of inorganic phosphate, was used as an expedient phosphoryl donor. The incipient αMan 1-P and guanosine triphosphate (GTP) were converted into GDP-Man by a highly manno compared to gluco selective nucleotidyltransferase. Pyrophosphatase was additionally required to hydrolyze the pyrophosphate released from the GTP, thus driving the reaction towards GDP-Man. The enzymatic cascade was operated with the αMan 1-P and the GDP-Man formation decoupled from one another (sequential mode) or having all steps run concurrently (simultaneous mode). Detailed time course analysis revealed that kinetic pull due to the constant removal of the intermediate αMan 1-P in simultaneous-mode reactions was important to promote phosphorylation of Man from αGlc 1-P in high efficiency, avoiding loss of sugar 1-phosphates by hydrolysis. Under optimized conditions for the one-pot transformation involving four enzymes, 100 mM (67 g L?1) GDP-Man was prepared from 140 mM sucrose and phosphate, using 400 mM Man as the phosphoryl acceptor. The product was recovered by anion-exchange and size-exclusion chromatography in ≥95% purity in about 50% yield (100 mg). These results demonstrate for the first time the practical use of a phosphorylase-phosphatase combi-catalyst as an alternative to the canonical kinase for the anomeric phosphorylation of the sugar substrate in nucleoside diphospho-sugar synthesis. Phosphorylation from inorganic phosphate via the intermediate αGlc 1-P rather than from NTP, particularly GTP, appears advantageous specifically in cases where the sugar acceptor is a bulk commodity that can be applied in suitable excess to the phosphatase reaction. (Figure presented.).
Studies on the substrate specificity of a GDP-mannose pyrophosphorylase from Salmonella enterica
Zou, Lu,Zheng, Ruixiang Blake,Lowary, Todd L.
supporting information; experimental part, p. 1219 - 1226 (2012/09/21)
A series of methoxy and deoxy derivatives of mannopyranose-1-phosphate (Manp-1P) were chemically synthesized, and their ability to be converted into the corresponding guanosine diphosphate mannopyranose (GDP-Manp) analogues by a pyrophosphorylase (GDP-ManPP) from Salmonella enterica was studied. Evaluation of methoxy analogues demonstrated that GDP-ManPP is intolerant of bulky substituents at the C-2, C-3, and C-4 positions, in turn suggesting that these positions are buried inside the enzyme active site. Additionally, both the 6-methoxy and 6-deoxy Manp-1P derivatives are good or moderate substrates for GDP-ManPP, thus indicating that the C-6 hydroxy group of the Manp-1P substrate is not required for binding to the enzyme. When taken into consideration with other previously published work, it appears that this enzyme has potential utility for the chemoenzymatic synthesis of GDP-Manp analogues, which are useful probes for studying enzymes that employ this sugar nucleotide as a substrate.
