@article{LIANG2022, 
author = {Shanquan LIANG and Dengya ZHANG and Shaoqing YANG and Zi HE and Dan LIU and Zhengqiang JIANG},
title = {Effect of Pathway Configurations on the Synthesis of 2'-Fucosyllactose in Escherichia coli},
year = {2022},
journal = {Food Science},
volume = {43},
number = {24},
pages = {110-116},
keywords = {biosynthesis, CRISPR/Cas9, Escherichia coli, 2'-fucosyllactose, pathway configurations},
url = {https://www.sciopen.com/article/10.7506/spkx1002-6630-20220602-016},
doi = {10.7506/spkx1002-6630-20220602-016},
abstract = {The de novo synthesis pathway of 2'-fucosyllactose (2'-FL) was established in Escherichia coli BL21 Star (DE3) in this study. The β-galactosidase gene lacZ M15 and the uridine diphosphate (UDP)-glucose lipid carrier transferase gene wcaJ were knocked out using the CRISPR/Cas9 gene editing system. The effects of three different pathway configurations, viz., operon, pseudo-operon, and monocistronic on the synthesis of 2'-FL were explored. The results showed that the concentration of 2'-FL produced in shake flasks was 0.34 g/L after overexpression of the de novo synthesis pathway related genes in E. coli BL21 Star (DE3). The concentration of 2'-FL was increased to 1.26 g/L by deleting the lacZ M15 and wcaJ genes. The highest concentration of 2'-FL of 1.92 g/L was observed in strain BS-7 when regulated by the operon expression. Fed-batch fermentation of strain BS-7 accumulated 14.04 g/L 2'-FL with a productivity of 0.59 g/(L·h) and a lactose conversion rate of 63%, respectively. This study suggested that lower gene expression levels not only increased 2'-FL production, but also could improve the conversion efficiency of substrate in engineered E. coli.}
}