Abstract: Guanosine 5′-diphospho-β-L-fucose (GDP-L-Fuc) is a key intermediate for fucosylated oligosaccharides biosynthesis in plants and bacteria. In this study, a gene encoding L-fucose kinase/GDP-L-fucose pyrophosphorylase (FKP) from Bacteroides fragilis was cloned into pET-22b(+) and expressed in Escherichia coli BL21(DE3). The purified recombinant FKP had the maximum activity at 45 ℃ and 7.5. The enzyme was stable at between pH 7.5 and 9.5 and at 4 ℃ and 12 ℃. Under the optimal reaction conditions, its Km and Kcat were 0.44 mmol/L and (31.49±2.5) s-1, respectively. With FKP catalyzing at 45 ℃ for 11 h, 2.1 mmol/L of GDP-L-fucose from 10 mmol/L of L-fucose was finally synthesized.
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2020, Vol. 46 
