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J Biotechnol. 2016 Oct 10;235:61-83. doi: 10.1016/j.jbiotec.2016.03.052. Epub 2016 Apr 1.

Biotechnological production of fucosylated human milk oligosaccharides: Prokaryotic fucosyltransferases and their use in biocatalytic cascades or whole cell conversion systems.

Author information

1
Graz University of Technology, Institute of Biotechnology and Biochemical Engineering, NAWI Graz, Petersgasse 12, A-8010 Graz, Austria; Austrian Centre of Industrial Biotechnology, Petersgasse 14, A-8010 Graz, Austria.
2
Graz University of Technology, Institute of Biotechnology and Biochemical Engineering, NAWI Graz, Petersgasse 12, A-8010 Graz, Austria; Austrian Centre of Industrial Biotechnology, Petersgasse 14, A-8010 Graz, Austria. Electronic address: bernd.nidetzky@tugraz.at.

Abstract

Human milk oligosaccharides (HMOs) constitute a class of complex carbohydrates unique to mother's milk and are strongly correlated to the health benefits of breastfeeding in infants. HMOs are important as functional ingredients of advanced infant formula and have attracted broad interest for use in health-related human nutrition. About 50% of the HMOs structures contain l-fucosyl residues, which are introduced into nascent oligosaccharides by enzymatic transfer from GDP-l-fucose. To overcome limitation in the current availability of fucosylated HMOs, biotechnological approaches for their production have been developed. Functional expression of the fucosyltransferase(s) and effective supply of GDP-l-fucose, respectively, are both bottlenecks of the biocatalytic routes of synthesis. Strategies of in vitro and in vivo production of fucosylated HMOs are reviewed here. Besides metabolic engineering for enhanced HMO production in whole cells, the focus is on the characteristics and the heterologous overexpression of prokaryotic α1,2- and α1,3/4-fucosyltransferases. Up to 20g/L of fucosylated HMOs were obtained in optimized production systems. Optimized expression enabled recovery of purified fucosyltransferases in a yield of up to 45mg/L culture for α1,2-fucosyltransferases and of up to 200mg protein/L culture for α1,3/4-fucosyltransferases.

KEYWORDS:

Fucosyllactose; Fucosyltransferases; Human milk oligosaccharides; Metabolic engineering; Multienzyme cascade; Whole cell systems

PMID:
27046065
DOI:
10.1016/j.jbiotec.2016.03.052
[Indexed for MEDLINE]

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