Synthesis of fucosylated lacto-N-tetraose using whole-cell biotransformation.
Fucosylated oligosaccharides present a predominant group of free oligosaccharides found in human milk. Here, a microbial conversion of lactose, D-glucose and L-fucose to fucosylated lacto-N-tetraose by growing Escherichia coli cultures is presented. The recombinant expression of genes encoding for the β1,3-N-acetylglucosaminyltransferase (LgtA) and the β1,3-galactosyltransferase (WbgO) enables the whole-cell biotransformation of lactose to lacto-N-tetraose. By the additional expression of a recombinant GDP-L-fucose salvage pathway together with a bacterial fucosyltransferase, lacto-N-tetraose is further converted into the respective fucosylated compounds. The expression of a gene encoding the α1,2-fucosyltransferase (FutC) in the lacto-N-tetraose producing E. coli strain led to the formation of lacto-N-fucopentaose I, whereas the expression of a gene encoding the α1,4-fucosyltransferase (FucT14) mainly led to the conversion of lacto-N-tetraose to lacto-N-difucohexaose II.
Institute of Microbiology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
Florian
F
Baumgärtner
Institute of Microbiology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
Lukas
L
Jurzitza
Bioorganic Chemistry, Institute of Chemistry, University of Hohenheim, Garbenstrasse 30, 70599 Stuttgart, Germany.
Jürgen
J
Conrad
Bioorganic Chemistry, Institute of Chemistry, University of Hohenheim, Garbenstrasse 30, 70599 Stuttgart, Germany.
Uwe
U
Beifuss
Institute of Microbiology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
Georg A
GA
Sprenger
Institute of Microbiology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany. Electronic address: imbca@imb.uni-stuttgart.de.
Christoph
C
Albermann
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism; D005647:Fucosyltransferases / Q000378:metabolism; D005700:Galactosyltransferases / Q000235:genetics / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007785:Lactose / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008895:Milk, Human / Q000378:metabolism; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry
D001426:Bacterial Proteins; D009844:Oligosaccharides; C056146:lacto-N-difucohexose II; C045442:lacto-N-fucopentaose I; C013084:lacto-N-neotetraose; D005647:Fucosyltransferases; D005700:Galactosyltransferases; D017351:N-Acetylglucosaminyltransferases; D007785:Lactose
Biotransformation; Fucosyltransferase; Human milk oligosaccharides; Lacto-N-difucohexaose II; Lacto-N-fucopentaose I; Lacto-N-tetraose