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Literature summary extracted from

  • Dobson, R.C.; Valegard, K.; Gerrard, J.A.
    The crystal structure of three site-directed mutants of Escherichia coli dihydrodipicolinate synthase: further evidence for a catalytic triad (2004), J. Mol. Biol., 338, 329-339.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.3.3.7 overexpression of wild-type and mutant enzymes in strain XL1-Blue Escherichia coli

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.3.3.7 purified recombinant wild-type and mutant enzymes, hanging or sitting drop vapour diffusion method, at 4°C and 21°C, 0.006 ml protein solutions: about 10 mg/ml protein, 1.8 M K2PO4, pH 10.0, N-octyl-beta-R-glucopyranoside 6% w/v, + 2 ml reservoir solution: 1.8 M K2PO4, pH 10.0, 3-4 days, X-ray diffraction structure determination and analysis at 2.35-2.5 A resolution Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
4.3.3.7 additional information detailed structural properties of the mutant enzymes based on the crystal structure models Escherichia coli
4.3.3.7 T44V site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme Escherichia coli
4.3.3.7 Y107F site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme Escherichia coli
4.3.3.7 Y133F site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.3.3.7 0.08
-
pyruvate mutant T44V, pH 8.0, 30°C Escherichia coli
4.3.3.7 0.09
-
(S)-aspartate 4-semialdehyde mutant T44V, pH 8.0, 30°C Escherichia coli
4.3.3.7 0.11
-
(S)-aspartate 4-semialdehyde wild-type enzyme, pH 8.0, 30°C Escherichia coli
4.3.3.7 0.16
-
pyruvate mutant Y107F, pH 8.0, 30°C Escherichia coli
4.3.3.7 0.26
-
pyruvate wild-type enzyme, pH 8.0, 30°C Escherichia coli
4.3.3.7 0.58
-
(S)-aspartate 4-semialdehyde mutant Y107F, pH 8.0, 30°C Escherichia coli
4.3.3.7 2.7
-
(S)-aspartate 4-semialdehyde mutant Y133F, pH 8.0, 30°C Escherichia coli
4.3.3.7 35
-
pyruvate mutant Y133F, pH 8.0, 30°C Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.3.3.7 (S)-aspartate 4-semialdehyde + pyruvate Escherichia coli branch point reaction in the biosynthesis of lysine dihydrodipicolinate + H2O
-
?

Organism

EC Number Organism UniProt Comment Textmining
4.3.3.7 Escherichia coli P0A6L2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.3.3.7 recombinant wild-type 5.8fold, recombinant mutant Y107F 17.6fold, recombinant mutant T44V 15.8fold, and recombinant mutant Y133F 18fold Escherichia coli

Reaction

EC Number Reaction Comment Organism Reaction ID
4.3.3.7 pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O ping-pong kinetic and catalytic mechanism involves a catalytic triad which consists of Tyr133, Thr44, and Tyr107, overview Escherichia coli

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.3.3.7 0.0034
-
purified mutant T44V Escherichia coli
4.3.3.7 0.008
-
purified mutant Y133F Escherichia coli
4.3.3.7 0.27
-
purified mutant Y107F Escherichia coli
4.3.3.7 1.81
-
purified wild-type enzyme Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.3.3.7 (S)-aspartate 4-semialdehyde + pyruvate
-
Escherichia coli dihydrodipicolinate + H2O
-
?
4.3.3.7 (S)-aspartate 4-semialdehyde + pyruvate branch point reaction in the biosynthesis of lysine Escherichia coli dihydrodipicolinate + H2O
-
?

Subunits

EC Number Subunits Comment Organism
4.3.3.7 tetramer homotetramer of (alpha/beta)8 barrels, each containing one active site, crystal structure Escherichia coli

Synonyms

EC Number Synonyms Comment Organism
4.3.3.7 DHDPS
-
Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.3.3.7 0.12
-
(S)-aspartate 4-semialdehyde mutant T44V, pH 8.0, 30°C Escherichia coli
4.3.3.7 0.7
-
(S)-aspartate 4-semialdehyde mutant Y133F, pH 8.0, 30°C Escherichia coli
4.3.3.7 10.8
-
(S)-aspartate 4-semialdehyde mutant Y107F, pH 8.0, 30°C Escherichia coli
4.3.3.7 124
-
(S)-aspartate 4-semialdehyde wild-type enzyme, pH 8.0, 30°C Escherichia coli