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Literature summary for 2.1.3.3 extracted from

  • Ngu, L.; Winters, J.N.; Nguyen, K.; Ramos, K.E.; DeLateur, N.A.; Makowski, L.; Whitford, P.C.; Ondrechen, M.J.; Beuning, P.J.
    Probing remote residues important for catalysis in Escherichia coli ornithine transcarbamoylase (2020), PLoS ONE, 15, e0228487 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.1.3.3

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) Tuner cells Escherichia coli

Protein Variants

Protein Variants Comment Organism
C273A the mutant shows 2.8 and 3.2fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
D140N the mutant shows 5.4 and 28fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
D231A the mutant shows 450 and 580fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
E299D the mutant shows 2.5 and 4.2fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
E299Q the mutant shows 110 and 51fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
H272L the mutant shows 310 and 120fold and decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
H272N the mutant shows 4.2 and 3.4fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
Q104L the mutant shows 5.9 and 10fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
R57A the mutant shows 57 and 44fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
S61A the mutant shows 6.1 and 1.8fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
Y160F the mutant shows 6.7 and 8.2fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
Y160S the mutant shows 1.5 and 14fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
Y229F the mutant shows 3.5 and 2.7fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli
Y229S the mutant shows 1.7 and 1.4fold decrease of activity with respect to carbamoyl phosphate and L-ornithine, respectively, compared to the wild type enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.15
-
 Carbamoyl phosphate mutant enzyme D231A, at pH 8.5 and 20°C Escherichia coli
0.22
-
 Carbamoyl phosphate mutant enzyme E299D, at pH 8.5 and 20°C Escherichia coli
0.26
-
 Carbamoyl phosphate mutant enzyme C273A, at pH 8.5 and 20°C Escherichia coli
0.27
-
 Carbamoyl phosphate wild type enzyme, at pH 8.5 and 20°C Escherichia coli
0.28
-
 Carbamoyl phosphate mutant enzyme R57A, at pH 8.5 and 20°C Escherichia coli
0.3
-
 Carbamoyl phosphate mutant enzyme S61A, at pH 8.5 and 20°C Escherichia coli
0.3
-
 Carbamoyl phosphate mutant enzyme Y160S, at pH 8.5 and 20°C Escherichia coli
0.32
-
 Carbamoyl phosphate mutant enzyme Y229S, at pH 8.5 and 20°C Escherichia coli
0.33
-
 L-ornithine mutant enzyme E299Q, at pH 8.5 and 20°C Escherichia coli
0.33
-
 Carbamoyl phosphate mutant enzyme Y229F, at pH 8.5 and 20°C Escherichia coli
0.34
-
 Carbamoyl phosphate mutant enzyme E299Q, at pH 8.5 and 20°C Escherichia coli
0.4
-
 L-ornithine mutant enzyme H272N, at pH 8.5 and 20°C Escherichia coli
0.4
-
 Carbamoyl phosphate mutant enzyme Q104L, at pH 8.5 and 20°C Escherichia coli
0.44
-
 L-ornithine mutant enzyme R57A, at pH 8.5 and 20°C Escherichia coli
0.45
-
 L-ornithine mutant enzyme Y229F, at pH 8.5 and 20°C Escherichia coli
0.5
-
 Carbamoyl phosphate mutant enzyme H272N, at pH 8.5 and 20°C Escherichia coli
0.53
-
 L-ornithine mutant enzyme S61A, at pH 8.5 and 20°C Escherichia coli
0.53
-
 Carbamoyl phosphate mutant enzyme Y160F, at pH 8.5 and 20°C Escherichia coli
0.53
-
 L-ornithine wild type enzyme, at pH 8.5 and 20°C Escherichia coli
0.59
-
 L-ornithine mutant enzyme D231A, at pH 8.5 and 20°C Escherichia coli
0.61
-
 L-ornithine mutant enzyme Y229S, at pH 8.5 and 20°C Escherichia coli
0.68
-
 L-ornithine mutant enzyme H272L, at pH 8.5 and 20°C Escherichia coli
0.73
-
 L-ornithine mutant enzyme C273A, at pH 8.5 and 20°C Escherichia coli
0.81
-
 L-ornithine mutant enzyme Q104L, at pH 8.5 and 20°C Escherichia coli
0.9
-
 L-ornithine mutant enzyme E299D, at pH 8.5 and 20°C Escherichia coli
0.93
-
 Carbamoyl phosphate mutant enzyme H272L, at pH 8.5 and 20°C Escherichia coli
1
-
 L-ornithine mutant enzyme Y160F, at pH 8.5 and 20°C Escherichia coli
1.4
-
 Carbamoyl phosphate mutant enzyme D140N, at pH 8.5 and 20°C Escherichia coli
7.3
-
 L-ornithine mutant enzyme Y160S, at pH 8.5 and 20°C Escherichia coli
17
-
 L-ornithine mutant enzyme D140N, at pH 8.5 and 20°C Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
110000
-
 gel filtration Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
carbamoyl phosphate + L-ornithine Escherichia coli
-
 phosphate + L-citrulline
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P04391
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap column chromatography and Superdex 200 gel filtration Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
carbamoyl phosphate + L-ornithine
-
 Escherichia coli phosphate + L-citrulline
-
 ?

Subunits

Subunits Comment Organism
homotrimer
-
 Escherichia coli

Synonyms

Synonyms Comment Organism
ornithine transcarbamoylase
-
 Escherichia coli
OTC
-
 Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
68
-
 melting temperature of the apoenzyme Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.48
-
 Carbamoyl phosphate mutant enzyme D231A, at pH 8.5 and 20°C Escherichia coli
0.7
-
 L-ornithine mutant enzyme D231A, at pH 8.5 and 20°C Escherichia coli
4.2
-
 Carbamoyl phosphate mutant enzyme H272L, at pH 8.5 and 20°C Escherichia coli
4.3
-
 L-ornithine mutant enzyme H272L, at pH 8.5 and 20°C Escherichia coli
4.6
-
 Carbamoyl phosphate mutant enzyme E299Q, at pH 8.5 and 20°C Escherichia coli
5.1
-
 L-ornithine mutant enzyme E299Q, at pH 8.5 and 20°C Escherichia coli
7.5
-
 Carbamoyl phosphate mutant enzyme R57A, at pH 8.5 and 20°C Escherichia coli
8.2
-
 L-ornithine mutant enzyme R57A, at pH 8.5 and 20°C Escherichia coli
52
-
 L-ornithine mutant enzyme Q104L, at pH 8.5 and 20°C Escherichia coli
74
-
 Carbamoyl phosphate mutant enzyme S61A, at pH 8.5 and 20°C Escherichia coli
88
-
 Carbamoyl phosphate mutant enzyme Q104L, at pH 8.5 and 20°C Escherichia coli
95
-
 L-ornithine mutant enzyme H272N, at pH 8.5 and 20°C Escherichia coli
110
-
 L-ornithine mutant enzyme Y160F, at pH 8.5 and 20°C Escherichia coli
120
-
 Carbamoyl phosphate mutant enzyme E299D, at pH 8.5 and 20°C Escherichia coli
120
-
 Carbamoyl phosphate mutant enzyme Y160F, at pH 8.5 and 20°C Escherichia coli
130
-
 L-ornithine mutant enzyme Y229F, at pH 8.5 and 20°C Escherichia coli
140
-
 Carbamoyl phosphate mutant enzyme C273A, at pH 8.5 and 20°C Escherichia coli
150
-
 Carbamoyl phosphate mutant enzyme Y229F, at pH 8.5 and 20°C Escherichia coli
170
-
 L-ornithine mutant enzyme C273A, at pH 8.5 and 20°C Escherichia coli
170
-
 L-ornithine mutant enzyme E299D, at pH 8.5 and 20°C Escherichia coli
180
-
 Carbamoyl phosphate mutant enzyme H272N, at pH 8.5 and 20°C Escherichia coli
240
-
 L-ornithine mutant enzyme S61A, at pH 8.5 and 20°C Escherichia coli
290
-
 Carbamoyl phosphate mutant enzyme Y229S, at pH 8.5 and 20°C Escherichia coli
300
-
 Carbamoyl phosphate mutant enzyme Y160S, at pH 8.5 and 20°C Escherichia coli
350
-
 L-ornithine mutant enzyme Y229S, at pH 8.5 and 20°C Escherichia coli
360
-
 Carbamoyl phosphate mutant enzyme D140N, at pH 8.5 and 20°C Escherichia coli
410
-
 L-ornithine mutant enzyme Y160S, at pH 8.5 and 20°C Escherichia coli
410
-
 Carbamoyl phosphate wild type enzyme, at pH 8.5 and 20°C Escherichia coli
440
-
 L-ornithine wild type enzyme, at pH 8.5 and 20°C Escherichia coli
460
-
 L-ornithine mutant enzyme D140N, at pH 8.5 and 20°C Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.4
-
 L-ornithine mutant enzyme D231A, at pH 8.5 and 20°C Escherichia coli
3.4
-
 Carbamoyl phosphate mutant enzyme D231A, at pH 8.5 and 20°C Escherichia coli
5
-
 Carbamoyl phosphate mutant enzyme H272L, at pH 8.5 and 20°C Escherichia coli
7.1
-
 L-ornithine mutant enzyme H272L, at pH 8.5 and 20°C Escherichia coli
14
-
 Carbamoyl phosphate mutant enzyme E299Q, at pH 8.5 and 20°C Escherichia coli
16
-
 L-ornithine mutant enzyme E299Q, at pH 8.5 and 20°C Escherichia coli
19
-
 L-ornithine mutant enzyme R57A, at pH 8.5 and 20°C Escherichia coli
27
-
 Carbamoyl phosphate mutant enzyme R57A, at pH 8.5 and 20°C Escherichia coli
30
-
 L-ornithine mutant enzyme D140N, at pH 8.5 and 20°C Escherichia coli
46
-
 L-ornithine mutant enzyme S61A, at pH 8.5 and 20°C Escherichia coli
61
-
 L-ornithine mutant enzyme Y160S, at pH 8.5 and 20°C Escherichia coli
80
-
 L-ornithine mutant enzyme Q104L, at pH 8.5 and 20°C Escherichia coli
100
-
 L-ornithine mutant enzyme Y160F, at pH 8.5 and 20°C Escherichia coli
200
-
 L-ornithine mutant enzyme E299D, at pH 8.5 and 20°C Escherichia coli
230
-
 Carbamoyl phosphate mutant enzyme Y160F, at pH 8.5 and 20°C Escherichia coli
240
-
 L-ornithine mutant enzyme H272N, at pH 8.5 and 20°C Escherichia coli
250
-
 Carbamoyl phosphate mutant enzyme S61A, at pH 8.5 and 20°C Escherichia coli
260
-
 L-ornithine mutant enzyme C273A, at pH 8.5 and 20°C Escherichia coli
260
-
 Carbamoyl phosphate mutant enzyme Q104L, at pH 8.5 and 20°C Escherichia coli
290
-
 Carbamoyl phosphate mutant enzyme D140N, at pH 8.5 and 20°C Escherichia coli
310
-
 L-ornithine mutant enzyme Y229F, at pH 8.5 and 20°C Escherichia coli
360
-
 Carbamoyl phosphate mutant enzyme H272N, at pH 8.5 and 20°C Escherichia coli
440
-
 Carbamoyl phosphate mutant enzyme Y229F, at pH 8.5 and 20°C Escherichia coli
560
-
 Carbamoyl phosphate mutant enzyme C273A, at pH 8.5 and 20°C Escherichia coli
590
-
 L-ornithine mutant enzyme Y229S, at pH 8.5 and 20°C Escherichia coli
620
-
 Carbamoyl phosphate mutant enzyme E299D, at pH 8.5 and 20°C Escherichia coli
830
-
 L-ornithine wild type enzyme, at pH 8.5 and 20°C Escherichia coli
920
-
 Carbamoyl phosphate mutant enzyme Y229S, at pH 8.5 and 20°C Escherichia coli
1000
-
 Carbamoyl phosphate mutant enzyme Y160S, at pH 8.5 and 20°C Escherichia coli
1500
-
 Carbamoyl phosphate wild type enzyme, at pH 8.5 and 20°C Escherichia coli