Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β- N -acetylhexosaminidases
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| 024 | 7 | 0 | |a 10.1007/s00253-015-6550-0 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6550-0 | ||
| 245 | 0 | 0 | |a Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β- N -acetylhexosaminidases |h [Elektronische Daten] |c [Christian Nyffenegger, Rune Nordvang, Birgitte Zeuner, Mateusz Łężyk, Elisabetta Difilippo, Madelon Logtenberg, Henk Schols, Anne Meyer, Jørn Mikkelsen] |
| 520 | 3 | |a This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. The enzyme-encoding genes were identified by functional screening of a soil-derived metagenomic library. The β-N-acetylhexosaminidases were expressed in Escherichia coli with an N-terminal His6-tag and were purified by nickel affinity chromatography. The sequence similarities of the enzymes with their respective closest homologues are 59% for HEX1 and 51% for HEX2 on the protein level. Both β-N-acetylhexosaminidases are classified into glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively. The enzymes are able to hydrolyze N-acetylchitooligosaccharides with a degree of polymerization of two, three, and four. The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8% based on the donor substrate. In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a Functional screening |2 nationallicence | |
| 690 | 7 | |a Protein expression |2 nationallicence | |
| 690 | 7 | |a Synthetic biology |2 nationallicence | |
| 690 | 7 | |a Chito-oligosaccharides |2 nationallicence | |
| 690 | 7 | |a Lacto- N -triose II |2 nationallicence | |
| 700 | 1 | |a Nyffenegger |D Christian |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | |
| 700 | 1 | |a Nordvang |D Rune |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | |
| 700 | 1 | |a Zeuner |D Birgitte |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | |
| 700 | 1 | |a Łężyk |D Mateusz |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | |
| 700 | 1 | |a Difilippo |D Elisabetta |u Laboratory of Food Chemistry, Wageningen University, Bornse Weilanden 9, 6708, Wageningen, The Netherlands |4 aut | |
| 700 | 1 | |a Logtenberg |D Madelon |u Laboratory of Food Chemistry, Wageningen University, Bornse Weilanden 9, 6708, Wageningen, The Netherlands |4 aut | |
| 700 | 1 | |a Schols |D Henk |u Laboratory of Food Chemistry, Wageningen University, Bornse Weilanden 9, 6708, Wageningen, The Netherlands |4 aut | |
| 700 | 1 | |a Meyer |D Anne |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | |
| 700 | 1 | |a Mikkelsen |D Jørn |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/19(2015-10-01), 7997-8009 |x 0175-7598 |q 99:19<7997 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6550-0 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-015-6550-0 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Nyffenegger |D Christian |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Nordvang |D Rune |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Zeuner |D Birgitte |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Łężyk |D Mateusz |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Difilippo |D Elisabetta |u Laboratory of Food Chemistry, Wageningen University, Bornse Weilanden 9, 6708, Wageningen, The Netherlands |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Logtenberg |D Madelon |u Laboratory of Food Chemistry, Wageningen University, Bornse Weilanden 9, 6708, Wageningen, The Netherlands |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Schols |D Henk |u Laboratory of Food Chemistry, Wageningen University, Bornse Weilanden 9, 6708, Wageningen, The Netherlands |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Meyer |D Anne |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Mikkelsen |D Jørn |u Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, Building 229, Søltofts Plads, 2800, Kgs. Lyngby, Denmark |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/19(2015-10-01), 7997-8009 |x 0175-7598 |q 99:19<7997 |1 2015 |2 99 |o 253 | ||