High-Level <i>De Novo</i> Biosynthesis of 2′-Fucosyllactose by Metabolically Engineered <i>Escherichia coli</i>
Yuanlin Liu, Yingying Zhu, Li Wan, Roulin Chen, Wenli Zhang, Wanmeng Mu
Journal of Agricultural and Food Chemistry
Abstract
2'-Fucosyllactose (2'-FL) is the most abundant oligosaccharide in human milk. In this study, a highly efficient biosynthetic route for 2'-FL production was designed <i>via</i> the <i>de novo</i> pathway of GDP-l-fucose using engineered <i>Escherichia coli</i> BL21(DE3). Specifically, plasmid-based strains with previously deleted <i>lacZ</i> and <i>wcaJ</i> were further reconstructed by introducing <i>de novo</i> pathway genes and α1,2-fucosyltransferase-encoding <i>wbgL</i> to realize 2'-FL synthesis. The 2'-FL titer was enhanced to 3.92 g/L by further introducing <i>rcsA</i> and <i>rcsB</i>. Subsequently, the additional <i>wbgL</i> expression cassette was chromosomally integrated into <i>recA</i> locus to strengthen fucosylation reaction and a strong constitutive promoter (P<sub>J23119</sub>) was used to replace the original promoters of <i>manC-manB</i> and <i>gmd-wcaG</i> to improve 2'-FL synthesis. The maximal 2'-FL titer reached 9.06 and 79.23 g/L in shake-flask and fed-batch cultivation, respectively. The 2'-FL productivity reached 1.45 g/L/h, showing remarkable production potential in large-scale industrial application.
Extracted Claims
3 claims extracted from this paper into the knowledge graph
2'-Fucosyllactose (2'-FL) showed productivity 1.45 g/L/h
“The 2'-FL productivity reached 1.45 g/L/h, showing remarkable production potential in large-scale industrial application.”
2'-Fucosyllactose (2'-FL) reached maximal titer 9.06 g/L in shake-flask and 79.23 g/L in fed-batch cultivation
“The maximal 2'-FL titer reached 9.06 and 79.23 g/L in shake-flask and fed-batch cultivation, respectively.”
2'-Fucosyllactose (2'-FL) is produced by engineered Escherichia coli BL21(DE3)
“The 2'-FL titer was enhanced to 3.92 g/L by further introducing rcsA and rcsB.”