Use of trans-sialidase and sialyltransferase for synthesis of si

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical

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435 72, 435 97, 435101, 435175, 435193, 435194, C12P 1944, C12P 1900, C12P 1918, C12N 1100

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054098173

ABSTRACT:
A single vessel cyclic synthesis process for preparation of a sialyl.alpha.2.fwdarw.3.beta.galactoside is disclosed. In accordance with this process, a sialyltransferase acceptor is sialylated in an aqueous reaction medium by an .alpha.(2,3)sialyl transferase and CMP-sialic acid to form a sialyl donor substrate and CMP. In the presence of the trans-sialidase of Trypanosoma crusi, that sialyl donor substrate provides a sialyl group for a trans-sialidase acceptor, thereby preparing the sialyl.alpha.2.fwdarw.3.beta.galactoside. The .alpha.(2,3)sialyltransferase acceptor is reformed upon trans-sialidation of the latter acceptor, and the sialyl donor substrate is reformed using the .alpha.(2,3)sialyltransferase and a CMP-sialic acid recycling system that combines CMP with sialic acid that is also present in the vessel. The K.sub.m /V.sub.max value for the .alpha.(2,3)sialyltransferase acceptor is less than one-tenth the value of K.sub.m /V.sub.max of the trans-sialidase acceptor for the .alpha.(2,3)sialyltransferase.

REFERENCES:
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Schenkman et al "Cell" 65(7):1117-1125 (1991).
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Schenkman et al "J. Exp Med" 175(2) 567-75 (1992).
Auge et al "Carbohydrate Res." 200 (1990)257-268.

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