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Literature summary for 4.3.1.1 extracted from

  • Puthan Veetil, V.; Raj, H.; Quax, W.J.; Janssen, D.B.; Poelarends, G.J.
    Site-directed mutagenesis, kinetic and inhibition studies of aspartate ammonia lyase from Bacillus sp. YM55-1 (2009), FEBS J., 276, 2994-3007.
    View publication on PubMed
Search for more literature for 4.3.1.1

Cloned(Commentary)

Cloned (Comment) Organism
His-tagged version expressed in Escherichia coli TOP10 Bacillus sp. YM55-1

Protein Variants

Protein Variants Comment Organism
H188A responsible for binding the amino group of the substrate Bacillus sp. YM55-1
H188K responsible for binding the amino group of the substrate Bacillus sp. YM55-1
H188Q responsible for binding the amino group of the substrate Bacillus sp. YM55-1
H188R responsible for binding the amino group of the substrate Bacillus sp. YM55-1
K324A responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1
K324D responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1
K324H responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1
K324R responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1
K324S responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1
K324V responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1
N142A responsible for binding the amino group of the substrate Bacillus sp. YM55-1
N142Q responsible for binding the amino group of the substrate Bacillus sp. YM55-1
N326A responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1
N326Q responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1
S140A implicated in binding the C4 carboxylate group of substrate Bacillus sp. YM55-1
S140G/T141G implicated in binding the C4 carboxylate group of substrate Bacillus sp. YM55-1
S140K implicated in binding the C4 carboxylate group of substrate Bacillus sp. YM55-1
S140K/T141K implicated in binding the C4 carboxylate group of substrate Bacillus sp. YM55-1
S140R implicated in binding the C4 carboxylate group of substrate Bacillus sp. YM55-1
T101A responsible for binding the amino group of the substrate Bacillus sp. YM55-1
T101S responsible for binding the amino group of the substrate Bacillus sp. YM55-1
T141A implicated in binding the C4 carboxylate group of substrate Bacillus sp. YM55-1
T141K implicated in binding the C4 carboxylate group of substrate Bacillus sp. YM55-1
T141R implicated in binding the C4 carboxylate group of substrate Bacillus sp. YM55-1
T141V implicated in binding the C4 carboxylate group of substrate Bacillus sp. YM55-1
T187A responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1
T187S responsible for binding the C1 carboxylate group of substrate Bacillus sp. YM55-1

Inhibitors

Inhibitors Comment Organism Structure
3-nitropropionate competitive Bacillus sp. YM55-1
D-malate competitive Bacillus sp. YM55-1

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 L-aspartate above 1 M, N142Q mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 1 M, N326A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 1 M, N326Q mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 1 M, T101A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 1 M, T101S mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 1 M, T187A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 400 mM, S140A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 400 mM, T141K mutant protein Bacillus sp. YM55-1
1.7
-
 L-aspartate T141A mutant protein Bacillus sp. YM55-1
15
-
 L-aspartate wild-type protein Bacillus sp. YM55-1
27
-
 L-aspartate H188A mutant protein Bacillus sp. YM55-1
163
-
 L-aspartate T187S mutant protein Bacillus sp. YM55-1

Organism

Organism UniProt Comment Textmining
Bacillus sp. YM55-1
-
-
-

Purification (Commentary)

Purification (Comment) Organism
immobilized metal ion affinity chromatography (Ni2+) Bacillus sp. YM55-1

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-aspartate
-
 Bacillus sp. YM55-1 fumarate + NH3
-
 r

Synonyms

Synonyms Comment Organism
aspartase
-
 Bacillus sp. YM55-1
aspartate ammonia lyase
-
 Bacillus sp. YM55-1

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 L-aspartate above 0.016 1/sec, N326Q mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 0.027 1/sec, T141K mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 0.12 1/sec, N326A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 0.38 1/sec, T101A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 0.43 1/sec, T187A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 0.88 1/sec, N142Q mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 34 1/sec, T101S mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate above 40 1/sec, S140A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, H188A, H188R and H188K mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, K324A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, K324D mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, K324H mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, K324R mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, K324S mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, K324V mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, N142A mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, S140G/T141G mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, S140K mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, S140K/T141K mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, S140R mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, T141R mutant protein Bacillus sp. YM55-1
additional information
-
 L-aspartate below 0.001 1/sec, T141V mutant protein Bacillus sp. YM55-1
0.3
-
 L-aspartate T141A mutant protein Bacillus sp. YM55-1
0.7
-
 L-aspartate H188A mutant protein Bacillus sp. YM55-1
40
-
 L-aspartate wild-type protein Bacillus sp. YM55-1
190
-
 L-aspartate T187S mutant protein Bacillus sp. YM55-1

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.01
-
 3-nitropropionate H188A mutant protein Bacillus sp. YM55-1
0.4
-
 D-malate H188A mutant protein Bacillus sp. YM55-1
0.5
-
 3-nitropropionate T141A mutant protein Bacillus sp. YM55-1
2
-
 3-nitropropionate wild-type protein Bacillus sp. YM55-1
48
-
 D-malate T141A mutant protein Bacillus sp. YM55-1
68
-
 D-malate wild-type protein Bacillus sp. YM55-1

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.000016
-
 L-aspartate N326Q mutant protein Bacillus sp. YM55-1
0.000068
-
 L-aspartate T141K mutant protein Bacillus sp. YM55-1
0.00012
-
 L-aspartate N326A mutant protein Bacillus sp. YM55-1
0.00038
-
 L-aspartate T101A mutant protein Bacillus sp. YM55-1
0.00043
-
 L-aspartate T187A mutant protein Bacillus sp. YM55-1
0.00088
-
 L-aspartate N142Q mutant protein Bacillus sp. YM55-1
0.026
-
 L-aspartate H188A mutant protein Bacillus sp. YM55-1
0.034
-
 L-aspartate T101S mutant protein Bacillus sp. YM55-1
0.1
-
 L-aspartate S140A mutant protein Bacillus sp. YM55-1
0.18
-
 L-aspartate T141A mutant protein Bacillus sp. YM55-1
1.2
-
 L-aspartate T187S mutant protein Bacillus sp. YM55-1
2.7
-
 L-aspartate wild-type protein Bacillus sp. YM55-1