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Literature summary for 4.2.1.47 extracted from

  • King, J.D.; Poon, K.K.; Webb, N.A.; Anderson, E.M.; McNally, D.J.; Brisson, J.R.; Messner, P.; Garavito, R.M.; Lam, J.S.
    The structural basis for catalytic function of GMD and RMD, two closely related enzymes from the GDP-D-rhamnose biosynthesis pathway (2009), FEBS J., 276, 2686-2700.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
His6-GMD expressed from the pQE-30-gmd vector Pseudomonas aeruginosa

Inhibitors

Inhibitors Comment Organism Structure
GDP-mannose
-
Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.01402
-
GDP-mannose pH 7.5, 37°C Pseudomonas aeruginosa

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q51366
-
-

Purification (Commentary)

Purification (Comment) Organism
by ultrasonication, centrifugation, on Ni2+-nitrilotriacetic acid resin and by anion exchange chromatography, to greater than 95% purity Pseudomonas aeruginosa

Storage Stability

Storage Stability Organism
-80°C, 25% glycerol, 1 year Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
GDP-4-dehydro-6-deoxy-D-mannose i.e., GDP-6-deoxy-D-lyxo-hexos-4-ulose Pseudomonas aeruginosa GDP-D-rhamnose
-
?
GDP-mannose is a bifunctional 4,6-dehydratase, and a stereospecific 4-reductase. A conserved amino acid side chain (Arg185) may be crucial for orienting substrate and cofactor Pseudomonas aeruginosa GDP-4-dehydro-6-deoxy-D-mannose + H2O
-
?

Synonyms

Synonyms Comment Organism
GDP-D-mannose-4,6-dehydratase
-
Pseudomonas aeruginosa
GMD
-
Pseudomonas aeruginosa

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25
-
His6-GMD loses activity slowly in the course of enzyme-substrate incubations, particularly at 37°C, but 4,6-dehydratase activity is still detectable after incubation for 16 h at 25°C Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
8.82
-
GDP-mannose pH 7.5, 37°C Pseudomonas aeruginosa

Cofactor

Cofactor Comment Organism Structure
NADPH addition of NADPH results in the gradual, His6-GMD-dependent reduction of GDP-6-deoxy-D-lyxo-hexos-4-ulose to GDP-D-rhamnose Pseudomonas aeruginosa

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.002859
-
GDP-mannose pH 7.5, 37°C Pseudomonas aeruginosa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
630
-
GDP-mannose pH 7.5, 37°C Pseudomonas aeruginosa