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Disaccharide Synthesis using E. coli UDP-glucose regeneration system  

Oh, Jeong-Seok (School of Chemical and Biomolecular Engineering, Georgia Institute of Technology)
Publication Information
KSBB Journal / v.23, no.6, 2008 , pp. 474-478 More about this Journal
Abstract
UDP-glucose regeneration system using metabolic engineeringis unique and efficient strategy for oligosaccharide synthesis. To exploit the efficient UDP-glucose regeneration system, we introduced four enzymes, which would be important in partitioning the flux of UDP regenerationsuch as UDP-glucose pyrophosphorylase, UDP-Kinase gene, UDP-galactose 4-epimerase, and $\beta$-1, 4-galactasyltrasnsferase, into E. coli AD202. To determine the optimal expression level for UDP-regeneration, LacNAc concentration was compared depending on IPTG concentration. 0.5 mM IPTG induction showed the higher oligosaccharides synthesis. Using metabolic engineering under optimal IPTG induction, LacNAc synthesis of AD202/pQNGLU increased until 16 h and showed the 1.34 mM. This concentration is 10 times higher than that of control strain at same reaction time. Lactose of AD202/pQNGLU showed the maximum synthesis of 0.39 mM at 16 h and showed the 2.6 times higher than that of control strain.
Keywords
Metabolic engineering UDP-glucose regeneration; Oligosaccharide synthesis; E. coli whole-cell biocatalytic synthesis;
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