Search for bacterial 1,2-fucosyltransferases for whole-cell biosynthesis of 2-fucosyllactose in recombinant Escherichia coli
Emine Seydametova, Jiwon Yu, Jonghyeok Shin, Yoonjung Park, Chakhee Kim, Hooyeon Kim, Seok Hyeon Yu, Yong-Cheol Park and Dae-Hyuk Kweon
Microbiological Research 222: 35-42 (2019.05)
Abstract
2-Fucosyllactose (2-FL) is the most abundant human milk oligosaccharide and is important for infant nutrition and health. Because 2-FL has potential as a functional ingredient in advanced infant formula and as a prebiotic in various foods, a cost-effective method for 2-FL production is desirable. 1,2-Fucosyltransferase (1,2-FT) is one of the key enzymes enabling the microbial biosynthesis of this complex sugar. However, the 1,2-FTs reported so far for the whole-cell biosynthesis of 2-FL originate from pathogens, posing a potential hurdle for approval as a food production method depending on countries. In this study, 10 1,2-FT genes from bacteria of biosafety level one were identified, and the main features of the deduced amino acid sequences were characterized. Four codon-optimized 1,2-FT genes were synthesized and introduced into Escherichia coli L M15 strain containing the plasmid pBCGW encoding guanosine 5-diphosphate-l-fucose biosynthetic enzymes. Among the four genes, 2-FL was produced only by the 1,2-FT from Thermosynechococcus elongatus (Te2FT). Bifidobacterium thermacidophilum 1,2-FT (Bt2FT) showed high expression but was not active in E. coli L M15. The other two 1,2-FTs were not expressed to a detectable level. During batch flask fermentation of Te2FT-expressing E. coli L M15 cells, 0.49 g/L 2-FL was obtained after 72 h of induction. This is comparable to the values previously reported for 1,2-FTs from Helicobacter pylori and Bacteroides fragilis.
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