Efficient production of uridine 5'-diphospho-N-acetylglucosamine by the combination of three recombinant enzymes and yeast cells.
Uridine 5'-diphospho N-acetylglucosamine (UDP-GlcNAc) is an important nucleotide sugar in the biochemistry of all living organisms, and it is an important substrate in the synthesis of oligosaccharides. In the present work, three bioactive enzymes, namely, glucokinase (YqgR), GlcNAc-phosphate mutase (Agm1), and N-acetylglucosamine-1-phosphate uridyltransferase (GlmU), were produced effectively as soluble form in recombinant Escherichia coli. These three enzymes and dried yeast together were used to construct a multistep enzymatic system, which could produce UDP-GlcNAc efficiently with N-acetylglucosamine (GlcNAc) as the substrate. After the optimization of various reaction conditions, 31.5 mMUDP-GlcNAc was produced from 50 mMGlcNAc and 50 mMUMP.
Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou, P.R. China.
Jiewen
J
Zhou
Limei
L
Fan
Peilian
P
Wei
Lei
L
Huang
Jin
J
Cai
Zhinan
Z
Xu
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005941:Glucokinase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007218:Industrial Microbiology / Q000379:methods*; D019751:Intramolecular Transferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D009713:Nucleotidyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism*
D011994:Recombinant Proteins; D014537:Uridine Diphosphate N-Acetylglucosamine; D005941:Glucokinase; D009713:Nucleotidyltransferases; C023226:UDPacetylglucosamine pyrophosphorylase; D019751:Intramolecular Transferases