Whole cell biosynthesis of a functional oligosaccharide, 2′-fucosyllactose, using engineered Escherichia coli

Abstract

Background: 2 0 -Fucosyllactose (2-FL) is a functional oligosaccharide present in human milk which protects against the infection of enteric pathogens. Because 2-FL can be synthesized through the enzymatic fucosylation of lactose with guanosine 5 0 -diphosphate (GDP)-L-fucose by α-1,2-fucosyltransferase (FucT2), an 2-FL producing Escherichia coli can be constructed through overexpressing genes coding for endogenous GDP-L-fucose biosynthetic enzymes and heterologous fucosyltransferase.

Enhanced production of 2′-fucosyllactose in engineered Escherichia coli BL21star(DE3) by modulation of lactose metabolism and fucosyltransferase

“…As mentioned above, most of previous studies employed the E. coli JM strains (E. coli K-12 derivatives) to produce 2-FL because it can take up lactose but cannot metabolize lactose as a carbon source. This unique phenotype seems to be due to complete deletion of the lac operon in the chromosome and introduction of the F' episome containing the modified lac operon (lacI q lacZ M15) (Baumgärtner et al, 2013;Drouillard et al, 2006;Lee et al, 2012).…”

Enhancing GDP-fucose production in recombinant Escherichia coli by metabolic pathway engineering

“…Chemical synthesis of fucose-containing chemicals could be achieved but often via tedious steps [7]. By comparison, enzymatic synthesis could overcome the disadvantages of chemical synthesis and has been considered as a powerful method to accomplish the synthesis process by a series of glycosyltransferases [8]. For the enzymatic fucosylation reaction, the availability of guanosine 5 -diphosphate (GDP)-fucose served as the indispensable donor substrate for fucosyltransferase is one of the crucial factors.…”

Synthesis of fucosylated lacto-N-tetraose using whole-cell biotransformation

“…The overall predominant oligosaccharides in human milk, found in previous studies which were dominated by milk from so-called secretor women are 2'-fucosyllactose (2'-FL), lacto-Nfucopentaose I (LNF I), lacto-N-difucohexaose I (LNDFH I), and lacto-N-tetraose (LNT). 7 Focusing on neutral HMOs, previous studies described chemical, enzymatic or whole-cell syntheses of fucosyllactoses [8][9][10][11][12][13][14] and core structures such as lacto-N-neotetraose (LNnT) and LNT. [15][16][17][18][19] Fucosylated derivatives of the core structure LNnT were synthesized in whole-cell cultivations, 10,11,20 while syntheses of fucosylated LNT structures so far have been only described from in vitro enzymatic conversions.…”