Efficient Biosynthesis of 2'-Fucosyllactose Using Glycerol and Lactose by Engineered Escherichia coli

Abstract

Abstract: 2'-fucosyllactose (2'-FL) is the most abundant component in human milk oligosaccharides (HMOs), and its biosynthesis has been a research hotspot. In this study, the de novo synthesis pathway of 2'-FL was established in E. coli BL21star(DE3). The β-galactosidase gene (lacZ) and the UDP-glucose lipid carrier transferase gene (wcaJ) were knocked out by the CRISPR/Cas9 system. Effect of various exogenous α-1,2-fucosyltransferases on the 2′-FL synthesis was investigated. Subsequently, the transcriptional level of pathway genes was regulated, and finally the fermentation condition was optimized. The results showed that the 2'-FL yield of 0.34 g/L was obtained in shake flask with the overexpression of de novo synthesis pathway genes in E. coli BL21star(DE3). The titer of 2'-FL was increased to 2.12 g/L after deletion of the lacZ and wcaJ genes. Expression of α-1,2-fucosyltransferase Wcfb from Bacteroides fragilis in shake flask resulted in the highest production of 4.12 g/L and no byproduct was produced. After optimization of the fermentation condition, the 2′-FL yields of 5.01 and 31.2 g/L were obtained in shake-flask and a 5 L fermenter, respectively.

Key words: 2'-fucosyllactose, CRISPR/Cas9, α-1,2-fucosyltransferase, biosynthesis, optimization of fermentation conditions

CLC Number:

TS201.2

Shan-Quan LIANG Zheng-Qiang Jiang Shaoqing Yang. Efficient Biosynthesis of 2'-Fucosyllactose Using Glycerol and Lactose by Engineered Escherichia coli[J]. FOOD SCIENCE.

References

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