title: Synthesis of GDP-mannose using coupling fermentation of recombinant Escherichia coli.
abstract: Glucokinase (glk), phosphomannomutase (manB), and mannose-1-phosphate guanylytransferase (manC) are needed for the biosynthesis of GDP-mannose. A recombinant E. coli strain over-expressing these three genes was constructed to produce guanosine 5'-diphosphate (GDP)-mannose, the donor of GDP-fucose, an essential substrate for synthesis of fucosyloligosaccharides. In addition, the glk, manB, and manC genes were individually cloned into the expression vector pET-22b (+) to construct three recombinant E. coli strains pET-glk, pET-manB and pET-manC, respectively. Fermentation of the recombinant strain BL21/pET-glk-manB-manC had a conversion rate of 23% from mannose to GDP-mannose under IPTG induction, while coupling fermentation of the three recombinant strains BL21/pET-glk, BL21/pET-manB, BL21/pET-manC resulted in a conversion rate of 33% under the same induction conditions.
authors.affiliation: Key Laboratory of Industrial Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, People's Republic of China.
authors.forename: Jia
authors.initials: J
authors.lastname: Honghong
authors.forename: Lu
authors.initials: L
authors.lastname: Fuping
authors.forename: Li
authors.initials: L
authors.lastname: Yu
authors.forename: Liu
authors.initials: L
authors.lastname: Xiaoguang
authors.forename: Liu
authors.initials: L
authors.lastname: Yihan
authors.forename: Wang
authors.initials: W
authors.lastname: Hongbin
authors.forename: Li
authors.initials: L
authors.lastname: Jing
authors.forename: Cao
authors.initials: C
authors.lastname: Yueting
mesh_terms: D001483:Base Sequence; D001709:Biotechnology; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D015870:Gene Expression; D005798:Genes, Bacterial; D005941:Glucokinase / Q000235:genetics / Q000378:metabolism; D006155:Guanosine Diphosphate Mannose / Q000096:biosynthesis*; D007700:Kinetics; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism; D017875:Phosphotransferases (Phosphomutases) / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic
chemical_list: D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D006155:Guanosine Diphosphate Mannose; D005941:Glucokinase; D009713:Nucleotidyltransferases; C021546:mannose-1-phosphate guanylyltransferase (GDP); D017875:Phosphotransferases (Phosphomutases); C052799:phosphomannomutase