Novel method for enzymatic synthesis of CMP-NeuAc.

Article abstract of DOI:10.1271/bbb.65.1736

A novel method for synthesizing CMP-NeuAc was established. We first confirmed that the putative neuA gene of Haemophilus influenzae, identified by its whole genome sequence project, indeed encodes CMP-NeuAc synthetase (EC 2.7.7.43). The enzyme requires CTP as a cytidylyl donor for cytidylylation of NeuAc. The enzyme was coupled with an enzymatic CTP-generating system from CMP and inorganic polyphosphate as a sole phospho-donor driven by the combination of polyphosphate kinase and CMP kinase, where phosphorylation of CMP is done by the combined activity expressed by both enzymes, and subsequent phosphorylation of CDP by polyphosphate kinase itself occurred efficiently. When CMP-NeuAc synthetase of H. influenzae, polyphosphate kinase, and CMP kinase were added to the reaction mixture containing equimolar concentrations (15 mM) of CMP and NeuAc, and polyphosphate (150 mM in terms of phosphate), CMP-NeuAc was synthesized up to 10 mM in 67% yield.

Full text of DOI:10.1271/bbb.65.1736

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Bioscience, Biotechnology, and Biochemistry
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Novel Method for Enzymatic Synthesis of CMP-NeuAc
Kazuya ISHIGEa, Tomoki HAMAMOTOa, Toshikazu SHIBAb & Toshitada NOGUCHIa
a Biochemicals Division, YAMASA Corporation
b Division of Molecular Chemistry, Graduate School of Engineering, Hokkaido University
Published online: 22 May 2014.
To cite this article: Kazuya ISHIGE, Tomoki HAMAMOTO, Toshikazu SHIBA & Toshitada NOGUCHI (2001) Novel Method
for Enzymatic Synthesis of CMP-NeuAc, Bioscience, Biotechnology, and Biochemistry, 65:8, 1736-1740, DOI: 10.1271/
bbb.65.1736
To link to this article: http://dx.doi.org/10.1271/bbb.65.1736
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