Pathway Optimization of 2′-Fucosyllactose Production in Engineered <i>Escherichia coli</i>
Wen Li, Yingying Zhu, Li Wan, Cuie Guang, Wanmeng Mu
Journal of Agricultural and Food Chemistry
Abstract
2'-Fucosyllactose (2'-FL), one of the most valuable oligosaccharides in human milk, is used as an emerging food ingredient in the nutraceutical and food industries due to its numerous health benefits. Herein, the <i>de novo</i> and salvage pathways for GDP-fucose synthesis were engineered and optimized in <i>Escherichia coli</i> BL21 (DE3) to improve the production of 2'-FL. The <i>de novo</i> pathway genes encoding phosphomannomutase (ManB), mannose-1-phosphate guanyltransferase (ManC), GDP-d-mannose-4,6-dehydratase (Gmd), and GDP-l-fucose synthase (WcaG) combined with the gene from the salvage pathway encoding fucose kinase/fucose-1-phosphate guanylyltransferase (Fkp) were reconstructed in two vectors to evaluate the GDP-fucose biosynthesis. Then, the <i>fucT2</i> gene, encoding α1,2-fucosyltransferase, was introduced into the GDP-fucose-overproducing strains to realize 2'-FL biosynthesis. Furthermore, the genes in bypass pathways, including <i>lacZ</i>, <i>fucI</i>, <i>fucK</i>, and <i>wcaJ</i>, were inactivated to improve 2'-FL production. In addition, the two GDP-fucose synthesis pathways, along with <i>fucT2</i>, were transcriptionally fine-tuned to efficiently increase 2'-FL production. The final metabolically engineered <i>E. coli</i> produced 2.62 and 14.1 g/L in shake-flask and fed-batch cultivations, respectively.
Extracted Claims
3 claims extracted from this paper into the knowledge graph
2'-Fucosyllactose (2'-FL) is used as an emerging food ingredient
“2'-Fucosyllactose (2'-FL), one of the most valuable oligosaccharides in human milk, is used as an emerging food ingredient in the nutraceutical and food industries due to its numerous health benefits.”
2'-FL produced 2.62 and 14.1 g/L
“The final metabolically engineered E. coli produced 2.62 and 14.1 g/L in shake-flask and fed-batch cultivations, respectively.”
GDP-fucose synthesis was engineered and optimized in Escherichia coli BL21 (DE3)
“The de novo and salvage pathways for GDP-fucose synthesis were engineered and optimized in Escherichia coli BL21 (DE3) to improve the production of 2'-FL.”