Bacterial synthesis of polysialic acid lactosides in recombinant Escherichia coli K-12

Bacterial polysialyltransferases (PSTs) are processive enzymes involved in the synthesis of polysialic capsular polysaccharides. They can also synthesize polysialic acid in vitro from disialylated and trisialylated lactoside acceptors, which are the carbohydrate moieties of GD3 and GT3 gangliosides,...

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Bibliographic Details
Published in:Glycobiology (Oxford) 2016-07, Vol.26 (7), p.723-731
Main Authors: Richard, Emeline, Buon, Laurine, Drouillard, Sophie, Fort, Sébastien, Priem, Bernard
Format: Article
Language:English
Subjects:
Biotechnology
Escherichia coli K12 - genetics
Gangliosides
Gene Expression Regulation, Enzymologic - genetics
Glycosides - biosynthesis
Glycosides - genetics
Glycosylation
Lactosylceramides
Life Sciences
Polysaccharides, Bacterial - biosynthesis
Polysaccharides, Bacterial - genetics
Recombinant Proteins - genetics
Sialic Acids - biosynthesis
Sialic Acids - genetics
Sialyltransferases - genetics
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Summary:Bacterial polysialyltransferases (PSTs) are processive enzymes involved in the synthesis of polysialic capsular polysaccharides. They can also synthesize polysialic acid in vitro from disialylated and trisialylated lactoside acceptors, which are the carbohydrate moieties of GD3 and GT3 gangliosides, respectively. Here, we engineered a non-pathogenic Escherichia coli strain that overexpresses recombinant sialyltransferases and sialic acid synthesis genes and can convert an exogenous lactoside into polysialyl lactosides. Several PSTs were assayed for their ability to synthesize polysialyl lactosides in the recombinant strains. Fed-batch cultures produced α-2,8 polysialic acid or alternate α-2,8-2,9 polysialic acid in quantities reaching several grams per liter. Bacterial culture in the presence of propargyl-β-lactoside as the exogenous acceptor led to the production of conjugatable polysaccharides by means of copper-assisted click chemistry.
ISSN:0959-6658
1460-2423
DOI:10.1093/glycob/cww027