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Literature summary extracted from

  • Westman, E.L.; McNally, D.J.; Charchoglyan, A.; Brewer, D.; Field, R.A.; Lam, J.S.
    Characterization of WbpB, WbpE, and WbpD and reconstitution of a pathway for the biosynthesis of UDP-2,3-diacetamido-2,3-dideoxy-D-mannuronic acid in Pseudomonas aeruginosa (2009), J. Biol. Chem., 284, 11854-11862.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.1.1.335 synthesis synthesis of preparative quantities of 2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid and 2,3-diacetamido-2,3-dideoxy-D-glucuronic acid by coupled reaction of enzyme WbpB and transaminase WpbE Pseudomonas aeruginosa

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.335 expression in Escherichia coli Pseudomonas aeruginosa
2.3.1.201 expressed in Escherichia coli Tuner(DE3) cells Pseudomonas aeruginosa
2.6.1.98 gene wbpE, expression of His6-tagged WbpE in Escherichia coli strain Rosetta(DE3) Pseudomonas aeruginosa

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.1.201 22700
-
 x * 22700, predicted from amino acid sequence Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.6.1.98 UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate + 2-oxoglutarate Pseudomonas aeruginosa transaminase WbpE product is UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate UDP-2-acetamido-2-deoxy-alpha-D-ribo-hex-3-uluronate + L-glutamate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.335 Pseudomonas aeruginosa G3XD23
-
-
2.3.1.201 Pseudomonas aeruginosa G3XD01
-
-
2.6.1.98 Pseudomonas aeruginosa
-
 serotype O5, gene wbpE
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.201 nickel-nitrilotriacetic acid-agarose column chromatography Pseudomonas aeruginosa
2.6.1.98 recombinant His6-tagged WbpE from Escherichia coli strain Rosetta(DE3) Pseudomonas aeruginosa

Storage Stability

EC Number Storage Stability Organism
1.1.1.335 -20°C, stable for at least 3 weeks Pseudomonas aeruginosa
2.6.1.98 -20°C, purified recombinant His6-tagged WbpE, 3 weeks, stable Pseudomonas aeruginosa
2.6.1.98 precipitation occurs if the protein is left at ambient temperature or subjected to excessive freeze/thaw cycles Pseudomonas aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.335 2-acetamido-2-deoxy-alpha-D-glucuronic acid + NAD+ + L-glutamate activity of enzyme His6-WbpB is detected only in presence of transaminase WbpE, and vice versa Pseudomonas aeruginosa UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronic acid + NADH + 2-ketoglutarate product of the coupled reaction of enzyme WbpB and transaminase WbpE ?
2.3.1.201 acetyl-CoA + UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate
-
 Pseudomonas aeruginosa CoA + UDP-2,3-diacetamido-2,3-dideoxy-alpha-D-glucuronate
-
 ?
2.6.1.98 additional information 2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid WbpE product identification by mass spectrometry, overview Pseudomonas aeruginosa ?
-
 ?
2.6.1.98 UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate + 2-oxoglutarate transaminase WbpE product is UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate Pseudomonas aeruginosa UDP-2-acetamido-2-deoxy-alpha-D-ribo-hex-3-uluronate + L-glutamate
-
 r
2.6.1.98 UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate + 2-oxoglutarate transaminase WbpE product is UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucuronate. Cofactor pyridoxal 5'-phosphate is converted to pyridoxamine phosphate during the reaction Pseudomonas aeruginosa UDP-2-acetamido-2-deoxy-alpha-D-ribo-hex-3-uluronate + L-glutamate
-
 r

Subunits

EC Number Subunits Comment Organism
2.3.1.201 ? x * 22700, predicted from amino acid sequence Pseudomonas aeruginosa

Synonyms

EC Number Synonyms Comment Organism
2.3.1.201 UDP-2-acetamido-3-amino-2,3-dideoxy-glucuronate N-acetyltransferase
-
 Pseudomonas aeruginosa
2.3.1.201 WbpD
-
 Pseudomonas aeruginosa
2.6.1.98 WbpE
-
 Pseudomonas aeruginosa

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.6.1.98 37
-
 assay at Pseudomonas aeruginosa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.6.1.98 7
-
 assay at Pseudomonas aeruginosa

Cofactor

EC Number Cofactor Comment Organism Structure
2.6.1.98 pyridoxal 5'-phosphate
-
 Pseudomonas aeruginosa

General Information

EC Number General Information Comment Organism
2.6.1.98 metabolism WbpE substrate 2-acetamido-2-deoxy-D-ribohex-3-uluronic acid is synthesized by WbpB Pseudomonas aeruginosa