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Literature summary extracted from

  • Lau, S.T.; Tanner, M.E.
    Mechanism and active site residues of GDP-fucose synthase (2008), J. Am. Chem. Soc., 130, 17593-17602.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.271 wild-type and mutants introduced into the pT7GFS vector. Plasmid encodes for the expression of GFS bearing N-terminal hexahistidine tags, expressed in Escherichia coli BL21 (DE3) Escherichia coli K-12

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.271 C109S active site mutant, produces GDP-6-deoxy-D-altrose as its major product (75%) besides GDP-L-fucose (25%) out of GDP-4-keto-6-deoxy-D-mannose Escherichia coli
1.1.1.271 C109S produces GDP-6-deoxy-D-altrose as its major product, thus C-3'' epimerization occurs first and premature reduction of the GDP-4-keto-6-deoxy-D-altrose intermediate becomes competitive with GDP-L-fucose production. Results in the appearance of a kinetic isotope effect when [3''-2H]-GDP-6-deoxy-4-ketomannose is used as a substrate. The mutant catalyzes a rapid wash-in of solvent derived deuterium into the C-5'' position of GDP-fucose in the presence of NADP+ Escherichia coli K-12
1.1.1.271 H179Q is inactive, readily catalyzes the wash-out of deuterium from the C-3'' position of [3''-2H]-GDP-6-deoxy-4-keto-mannose Escherichia coli K-12
1.1.1.271 H179Q the mutant catalyzes isotope exchange into starting material indicates that its active site has not been dramatically perturbed and it is still able to bind substrate and catalyze deprotonation events Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.271 Escherichia coli P32055
-
-
1.1.1.271 Escherichia coli K-12
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.271 by metal affinity chromatography Escherichia coli K-12

Storage Stability

EC Number Storage Stability Organism
1.1.1.271 -80°C, 3 months Escherichia coli K-12

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.271 GDP-4-keto-6-deoxy-D-mannose + NADPH + H+
-
 Escherichia coli K-12 GDP-L-fucose + NADP+
-
 ?
1.1.1.271 GDP-4-keto-6-deoxy-D-mannose + NADPH + H+
-
 Escherichia coli GDP-L-fucose + NADP+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.271 GDP-4-keto-6-deoxy-D-mannose epimerase/reductase
-
 Escherichia coli K-12
1.1.1.271 GDP-4-keto-6-deoxy-D-mannose epimerase/reductase
-
 Escherichia coli
1.1.1.271 GDP-fucose synthase
-
 Escherichia coli K-12
1.1.1.271 GDP-fucose synthase
-
 Escherichia coli
1.1.1.271 GFS
-
 Escherichia coli K-12
1.1.1.271 GFS
-
 Escherichia coli
1.1.1.271 GMER
-
 Escherichia coli K-12
1.1.1.271 GMER
-
 Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.271 1
-
 GDP-6-deoxy-4-keto-mannose mutant C109S, isotope [5''-2H]-GDP-6-deoxy-4-keto-mannose Escherichia coli K-12
1.1.1.271 1
-
 GDP-6-deoxy-4-keto-mannose wild-type, isotope [3''-2H]-GDP-6-deoxy-4-keto-mannose Escherichia coli K-12
1.1.1.271 1
-
 GDP-6-deoxy-4-keto-mannose wild-type, isotope [5''-2H]-GDP-6-deoxy-4-keto-mannose Escherichia coli K-12
1.1.1.271 1.5
-
 GDP-6-deoxy-4-keto-mannose mutant C109S, isotope [3''-2H]-GDP-6-deoxy-4-keto-mannose Escherichia coli K-12

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.271 NADPH
-
 Escherichia coli K-12
1.1.1.271 NADPH
-
 Escherichia coli