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Enzymatic synthesis and activation of sialic acid analogs

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Research objectives and content
The main objective of this research is to investigate the NeuAc system. This would require the parallel subcloning of two plasmid-based E. coli genes for synthesis (N-acetyl neuraminic acid synthase = NeuS; EC 4.1.3.19) and activation (CMP-N-acetyl neuraminic acid synthetase = CMP-NeuS; EC 2.7.7.43) of neuraminic acid analogs into a commercial expression vector under the control of a strong inducible promoter (e.g. pKK223-3; Pharmacia), as well as the overexpression, purification, and characterization of the corresponding enzymes for preparative applications. Since only few ManNAc analogs are commercially available for the synthesis of sialic acid analogs the first step is to synthesize selected ManNAc derivatives or analogs. In this sense, it would be interesting the synthesis of the thio derivative: that can be very useful in the study of the NeuAc system and in the synthesis of 6-thio-N-acetyl-Dneuraminic acid derivatives.
Training content (objective, benefit and expected impact)
The synthesis of sialic acids analogues presents an important problem in view of the interesting biological properties that can be anticipated. As well as the synthetic and analytical techniques, with which I have become familiarized during my PhD, this work would include basic techniques of gene technology, protein chemistry, and enzyme-catalyzed synthesis of nucleotide-activated sugars and oligosaccharides, allowing me to use a wide range of methods.

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Finanzierungsplan

RGI - Research grants (individual fellowships)

Koordinator

AACHEN UNIVERSITY OF TECHNOLOGY
EU-Beitrag
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Professor-Pirlet-straße 1
52074 AACHEN
Deutschland
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