Development of fluorescent Escherichia coli for a whole-cell sensor of 2ʹ-fucosyllactose

Jonghyeok Shin, Myungseo Park, Chakhee Kim, Hooyeon Kim, Yunjeong Park, Choongjin Ban, Jong Won Yoon, Chul Soo Shin, Jae Won Lee, Yong Su Jin, Yong Cheol Park, Won Ki Min, Dae Hyuk Kweon

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Abstract

2′-Fucosyllactose (2′-FL), a major component of fucosylated human milk oligosaccharides, is beneficial to human health in various ways like prebiotic effect, protection from pathogens, anti-inflammatory activity and reduction of the risk of neurodegeneration. Here, a whole-cell fluorescence biosensor for 2′-FL was developed. Escherichia coli (E. coli) was engineered to catalyse the cleavage of 2′-FL into l-fucose and lactose by constitutively expressing α-l-fucosidase. Escherichia coli ∆L YA, in which lacZ is deleted and lacY is retained, was employed to disable lactose consumption. E. coli ∆L YA constitutively co-expressing α-l-fucosidase and a red fluorescence protein (RFP) exhibited increased fluorescence intensity in media containing 2′-FL. However, the presence of 50 g/L lactose reduced the RFP intensity due to lactose-induced cytotoxicity. Preadaptation of bacterial strains to fucose alleviated growth hindrance by lactose and partially recovered the fluorescence intensity. The fluorescence intensity of the cell was linearly proportional to 1–5 g/L 2′-FL. The whole-cell sensor will be versatile in developing a 2′-FL detection system.

Original languageEnglish (US)
Article number10514
JournalScientific reports
Volume10
Issue number1
DOIs
StatePublished - Dec 1 2020

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    Shin, J., Park, M., Kim, C., Kim, H., Park, Y., Ban, C., Yoon, J. W., Shin, C. S., Lee, J. W., Jin, Y. S., Park, Y. C., Min, W. K., & Kweon, D. H. (2020). Development of fluorescent Escherichia coli for a whole-cell sensor of 2ʹ-fucosyllactose. Scientific reports, 10(1), [10514]. https://doi.org/10.1038/s41598-020-67359-x