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Preparative Enzymatic Synthesis of Activated Neuraminic Acid by Using a Microbial Enzyme

1992; Wiley; Volume: 672; Issue: 1 Enzyme Engine Linguagem: Inglês

10.1111/j.1749-6632.1992.tb32711.x

1749-6632

Matthias Kittelmann, T. KLEIN, Udo Kragl, Christian Wandrey, Oreste Ghisalba,

Carbohydrate Chemistry and Synthesis

Using Escherichia coli K-235 as a production strain in a fed-batch fermentation process with an optimized sorbitol/yeast extract medium, we were able to produce 640 U of CMP-Neu5Ac synthetase in 10 l scale (64 U/l) and 9200 U (total enzyme) in 200 l scale (390 U/kg wet weight). By simple one-step purification procedures, enzyme preparations were obtained that could be used efficiently for the synthesis of CMP-Neu5Ac from CTP and Neu5Ac with over 90% yield, from Neu5Ac, CMP, and ATP or phosphoenolpyruvate by in situ generation of CTP, and from CTP, pyruvate, and ManNAc or GlcNAc by in situ generation of Neu5Ac.