Sie befinden Sich nicht im Netzwerk der Universität Paderborn. Der Zugriff auf elektronische Ressourcen ist gegebenenfalls nur via VPN oder Shibboleth (DFN-AAI) möglich. mehr Informationen...
Synthesis of the Human Milk Oligosaccharide Lacto-N-Tetraose in Metabolically Engineered, Plasmid-Free E. coli
Ist Teil von
Chembiochem : a European journal of chemical biology, 2014-09, Vol.15 (13), p.1896-1900
Ort / Verlag
Weinheim: WILEY-VCH Verlag
Erscheinungsjahr
2014
Link zum Volltext
Quelle
Wiley Online Library - AutoHoldings Journals
Beschreibungen/Notizen
Human milk oligosaccharides (HMOs) constitute the third most abundant solid component of human milk. HMOs have been demonstrated to show positive effects on the infant’s well‐being. Despite numerous studies, more physiological analyses of single compounds are needed to fully elucidate these effects. Although being one of the most abundant core structures in human milk, the HMO lacto‐N‐tetraose (LNT) is not available at reasonable prices. In this study, we demonstrate the construction of the first E. coli strain capable of producing LNT in vivo. The strain was constructed by chromosomally integrating the genes lgtA and wbgO, encoding β‐1,3‐N‐acetylglucosaminyltransferase and β‐1,3‐galactosyltransferase. In shake‐flask cultivations, the strain yielded a total concentration of 219.1±3.5 mg L−1 LNT (LNT in culture broth and the cell pellet). After recovery of LNT, structural analysis by NMR spectroscopy confirmed the molecule structure.
Formula milk: Construction of the first E. coli strain capable of in vivo synthesis of the human milk oligosaccharide lacto‐N‐tetraose is reported. Chromosomal integration of recombinant genes encoding bacterial glycosyltransferases LgtA and WbgO allows cellular synthesis in shake flasks from glucose/glycerol (bulk carbon sources) and lactose. Extensive structural analysis unambiguously confirmed the structure of lacto‐N‐tetraose.