Large-scale production of UDP-galactose and globotriose by coupling metabolically engineered bacteria

• Published in: Nature biotechnology Vol. 16; no. 9; pp. 847 - 850
• Main Authors: Koizumi, Satoshi, Endo, Tetsuo, Tabata, Kazuhiko, Ozaki, Akio
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.09.1998
• Subjects: Base Sequence; Bioconversions. Hemisynthesis; Biological and medical sciences; Biotechnology; Carbohydrate Conformation; Carbohydrate Sequence; Cloning, Molecular; Corynebacterium - genetics; DNA Primers; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Methods. Procedures. Technologies; Molecular Sequence Data; Neisseria gonorrhoeae - genetics; Plasmids; Recombination, Genetic; Trisaccharides - biosynthesis; Trisaccharides - chemistry; Uridine Diphosphate Galactose - biosynthesis; Recombinant microorganism; Enzyme; Transferases; Escherichia coli; Oligosaccharide; Glycosyltransferases; Biosynthesis; Gene; Production; Bacteria; Actinomycetes; Corynebacteriaceae; Trisaccharide; Enterobacteriaceae; Ose nucleotide
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt0998-847

A large-scale production system of uridine 5'-diphospho-galactose (UDP-Gal) has been established by the combination of recombinant Escherichia coli and Corynebacterium ammoniagenes. Recombinant E. coli that overexpress the UDP-Gal biosynthetic genes galT, galK, and galU were generated. C. ammoniagenes contribute the production of uridine triphosphate (UTP), a substrate for UDP-Gal biosynthesis, from orotic acid, an inexpensive precursor of UTP. UDP-Gal accumulated to 72 mM (44 g/L) after a 21 h reaction starting with orotic acid and galactose. When E. coli cells that expressed the alpha1,4-galactosyltransferase gene of Neisseria gonorrhoeae were coupled with this UDP-Gal production system, 372 mM (188 g/L) globotriose (Galalpha1-4Galbeta1-4Glc), a trisaccharide portion of verotoxin receptor, was produced after a 36 h reaction starting with orotic acid, galactose, and lactose. No oligosaccharide by-products were observed in the reaction mixture. The production of globotriose was several times higher than that of UDP-Gal. The strategy of producing sugar nucleotides by combining metabolically engineered recombinant E. coli with a nucleoside 5'-triphosphate producing microorganism, and the concept of producing oligosaccharides by coupling sugar nucleotide production systems with glycosyltransferases, can be applied to the manufacture of other sugar nucleotides and oligosaccharides.