BRENDA - Enzyme Database show
show all sequences of 4.3.1.2

The roles of active site residues in the catalytic mechanism of methylaspartate ammonia-lyase

Raj, H.; Poelarends, G.J.; FEBS open bio 3, 285-290 (2013)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli
Clostridium tetanomorphum
Engineering
Amino acid exchange
Commentary
Organism
C361A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 47% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
C361K
mutant shows no reactivity. Conversion 2h: 10% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
F170A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 63% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
L384A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 61% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
Q172A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 16% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
Q172N
mutant shows no reactivity. Conversion 2h: 6% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
Q73A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 37% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 91:9 (89:11 wild-type)
Clostridium tetanomorphum
Q73N
mutant shows very low-level amination activity preventing the measurement of kinetic parameters. Conversion 2h: 18% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
T360A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 38% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 93:7 (89:11 wild-type)
Clostridium tetanomorphum
T360S
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 76% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 86:14 (89:11 wild-type)
Clostridium tetanomorphum
Y356A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 24% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.2
-
mesaconate
pH 9, 30°C, mutant Q73A
Clostridium tetanomorphum
0.7
-
mesaconate
pH 9, 30°C, wild-type
Clostridium tetanomorphum
0.9
-
mesaconate
pH 9, 30°C, mutant F170A
Clostridium tetanomorphum
1.4
-
mesaconate
pH 9, 30°C, mutant T360S
Clostridium tetanomorphum
2.9
-
mesaconate
pH 9, 30°C, mutant C361A
Clostridium tetanomorphum
15
-
mesaconate
pH 9, 30°C, mutant T360A
Clostridium tetanomorphum
60
-
mesaconate
pH 9, 30°C, mutant L384A; pH 9, 30°C, mutant Q172A; pH 9, 30°C, mutant Y356A
Clostridium tetanomorphum
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Clostridium tetanomorphum
Q05514
-
-
Purification (Commentary)
Commentary
Organism
-
Clostridium tetanomorphum
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
mesaconate + NH3
conversion 2h: 73% (wild-type), diastereomeric product ratio (threo:erythro) after 2h: 89:11
729745
Clostridium tetanomorphum
L-threo-3-methylaspartate + L-erythro-3-methylaspartate
-
-
-
r
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Clostridium tetanomorphum
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.6
-
mesaconate
pH 9, 30°C, mutant T360A
Clostridium tetanomorphum
0.7
-
mesaconate
pH 9, 30°C, mutant Q73A; pH 9, 30°C, mutant Y356A
Clostridium tetanomorphum
10
-
mesaconate
pH 9, 30°C, mutant F170A
Clostridium tetanomorphum
16
-
mesaconate
pH 9, 30°C, mutant L384A
Clostridium tetanomorphum
20
-
mesaconate
pH 9, 30°C, mutant Q172A
Clostridium tetanomorphum
27.5
-
mesaconate
pH 9, 30°C, mutant C361A
Clostridium tetanomorphum
35
-
mesaconate
pH 9, 30°C, mutant T360S
Clostridium tetanomorphum
61
-
mesaconate
pH 9, 30°C, wild-type
Clostridium tetanomorphum
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9
-
assay at
Clostridium tetanomorphum
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli
Clostridium tetanomorphum
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
C361A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 47% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
C361K
mutant shows no reactivity. Conversion 2h: 10% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
F170A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 63% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
L384A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 61% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
Q172A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 16% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
Q172N
mutant shows no reactivity. Conversion 2h: 6% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
Q73A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 37% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 91:9 (89:11 wild-type)
Clostridium tetanomorphum
Q73N
mutant shows very low-level amination activity preventing the measurement of kinetic parameters. Conversion 2h: 18% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
T360A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 38% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 93:7 (89:11 wild-type)
Clostridium tetanomorphum
T360S
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 76% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 86:14 (89:11 wild-type)
Clostridium tetanomorphum
Y356A
kcat (mesaconate) decreased compared to wild-type, Km (mesaconate) increased compared to wild-type. Conversion 2h: 24% (73% wild-type), diastereomeric product ratio (threo:erythro) after 2h: 95:5 (89:11 wild-type)
Clostridium tetanomorphum
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.2
-
mesaconate
pH 9, 30°C, mutant Q73A
Clostridium tetanomorphum
0.7
-
mesaconate
pH 9, 30°C, wild-type
Clostridium tetanomorphum
0.9
-
mesaconate
pH 9, 30°C, mutant F170A
Clostridium tetanomorphum
1.4
-
mesaconate
pH 9, 30°C, mutant T360S
Clostridium tetanomorphum
2.9
-
mesaconate
pH 9, 30°C, mutant C361A
Clostridium tetanomorphum
15
-
mesaconate
pH 9, 30°C, mutant T360A
Clostridium tetanomorphum
60
-
mesaconate
pH 9, 30°C, mutant L384A; pH 9, 30°C, mutant Q172A; pH 9, 30°C, mutant Y356A
Clostridium tetanomorphum
Purification (Commentary) (protein specific)
Commentary
Organism
-
Clostridium tetanomorphum
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
mesaconate + NH3
conversion 2h: 73% (wild-type), diastereomeric product ratio (threo:erythro) after 2h: 89:11
729745
Clostridium tetanomorphum
L-threo-3-methylaspartate + L-erythro-3-methylaspartate
-
-
-
r
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Clostridium tetanomorphum
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.6
-
mesaconate
pH 9, 30°C, mutant T360A
Clostridium tetanomorphum
0.7
-
mesaconate
pH 9, 30°C, mutant Q73A; pH 9, 30°C, mutant Y356A
Clostridium tetanomorphum
10
-
mesaconate
pH 9, 30°C, mutant F170A
Clostridium tetanomorphum
16
-
mesaconate
pH 9, 30°C, mutant L384A
Clostridium tetanomorphum
20
-
mesaconate
pH 9, 30°C, mutant Q172A
Clostridium tetanomorphum
27.5
-
mesaconate
pH 9, 30°C, mutant C361A
Clostridium tetanomorphum
35
-
mesaconate
pH 9, 30°C, mutant T360S
Clostridium tetanomorphum
61
-
mesaconate
pH 9, 30°C, wild-type
Clostridium tetanomorphum
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9
-
assay at
Clostridium tetanomorphum
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.04
-
mesaconate
pH 9, 30°C, mutant T360A
Clostridium tetanomorphum
0.27
-
mesaconate
pH 9, 30°C, mutant L384A
Clostridium tetanomorphum
0.342
-
mesaconate
pH 9, 30°C, mutant Q172A
Clostridium tetanomorphum
3.5
-
mesaconate
pH 9, 30°C, mutant Q73A
Clostridium tetanomorphum
9.5
-
mesaconate
pH 9, 30°C, mutant C361A
Clostridium tetanomorphum
11
-
mesaconate
pH 9, 30°C, mutant F170A
Clostridium tetanomorphum
12
-
mesaconate
pH 9, 30°C, mutant Y356A
Clostridium tetanomorphum
25
-
mesaconate
pH 9, 30°C, mutant T360S
Clostridium tetanomorphum
87
-
mesaconate
pH 9, 30°C, wild-type
Clostridium tetanomorphum
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.04
-
mesaconate
pH 9, 30°C, mutant T360A
Clostridium tetanomorphum
0.27
-
mesaconate
pH 9, 30°C, mutant L384A
Clostridium tetanomorphum
0.342
-
mesaconate
pH 9, 30°C, mutant Q172A
Clostridium tetanomorphum
3.5
-
mesaconate
pH 9, 30°C, mutant Q73A
Clostridium tetanomorphum
9.5
-
mesaconate
pH 9, 30°C, mutant C361A
Clostridium tetanomorphum
11
-
mesaconate
pH 9, 30°C, mutant F170A
Clostridium tetanomorphum
12
-
mesaconate
pH 9, 30°C, mutant Y356A
Clostridium tetanomorphum
25
-
mesaconate
pH 9, 30°C, mutant T360S
Clostridium tetanomorphum
87
-
mesaconate
pH 9, 30°C, wild-type
Clostridium tetanomorphum
Other publictions for EC 4.3.1.2
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
747577
Abidin
Modular enzymatic cascade syn ...
Clostridium tetanomorphum
Chemistry
24
17434-17438
2018
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1
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1
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3
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3
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1
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3
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1
1
-
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-
729599
PuthanVeetil
-
Enantioselective Synthesis of ...
Clostridium tetanomorphum
ChemCatChem
5
1325-1327
2013
-
-
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1
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1
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14
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1
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1
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1
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14
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1
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1
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729745
Raj
The roles of active site resid ...
Clostridium tetanomorphum
FEBS open bio
3
285-290
2013
-
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1
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11
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7
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2
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1
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1
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1
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11
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8
1
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9
9
729057
Raj
Characterization of a thermost ...
Carboxydothermus hydrogenoformans
Appl. Microbiol. Biotechnol.
94
385-397
2012
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1
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2
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2
1
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5
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11
1
1
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2
1
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1
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11
1
1
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1
2
1
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2
2
730442
Raj
Engineering methylaspartate am ...
Clostridium tetanomorphum
Nat. Chem.
4
478-484
2012
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1
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2
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1
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1
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35
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1
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1
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1
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35
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1
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1
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714983
Ramezani
Glutamate racemization and cat ...
Fusobacterium varium
FEBS J.
278
2540-2551
2011
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1
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703197
Raj
Alteration of the diastereosel ...
Clostridium tetanomorphum
ChemBioChem
10
2236-2245
2009
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1
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3
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5
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1
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1
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6
1
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1
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5
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2
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1
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6
1
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8
8
682798
Potrykus
Proteomic investigation of glu ...
Fusobacterium varium
Proteomics
7
1839-1853
2007
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1
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1
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1
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1
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1
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682889
Lee
Synthesis of mono- and di-deut ...
Escherichia coli
Tetrahedron
63
4663-4668
2007
-
1
1
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1
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1
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1
1
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1
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663816
Asano
-
Structure and Function of Amin ...
Citrobacter amalonaticus, Citrobacter amalonaticus YG-1002, Citrobacter freundii, Citrobacter freundii YG.0504, Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920, Morganella morganii, Morganella morganii YG-0601
Biocatal. Biotransform.
22
131-138
2004
-
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1
1
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3
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6
6
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23
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6
3
1
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2
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1
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1
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3
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6
6
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6
3
1
-
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2
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652354
Asuncion
The structure of 3-methylaspar ...
Clostridium tetanomorphum
J. Biol. Chem.
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8306-8311
2002
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653894
Levy
Insights into enzyme evolution ...
Citrobacter amalonaticus
Structure
10
105-113
2002
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649166
Asuncion
Overexpression, purification, ...
Clostridium tetanomorphum
Acta Crystallogr. Sect. D
57
731-733
2001
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653208
Bearne
Purification and characterizat ...
Fusobacterium varium
Mol. Cell. Biochem.
221
117-126
2001
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6064
Gani
The methylaspartase reaction p ...
Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920
Bioorg. Med. Chem.
7
977-990
1999
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650542
Pollard
Mechanism of 3-methylaspartase ...
Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920
Bioorg. Med. Chem.
7
949-975
1999
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6062
Kato
Cloning, nucleotide sequencing ...
Citrobacter amalonaticus, Citrobacter amalonaticus TPU 6323, Citrobacter amalonaticus YG-1002, Clostridium tetanomorphum, Clostridium tetanomorphum NCIMB 11547
Appl. Microbiol. Biotechnol.
50
468-474
1998
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5670
Kato
3-Methylaspartate ammonia-lyas ...
Citrobacter amalonaticus, Citrobacter amalonaticus YG-1002, Citrobacter freundii, Citrobacter freundii TPU 0011, Citrobacter freundii YG-0504, Citrobacter koseri, Citrobacter koseri JCM 1658, Citrobacter koseri JCM 1659, Clostridium tetanomorphum, Morganella morganii, Morganella morganii NCIMB 10466, Morganella morganii NCIMB 232, Morganella morganii NCIMB 9525, Morganella morganii YG-0601, Raoultella planticola, Raoultella planticola IAM 1133
Arch. Microbiol.
168
457-463
1997
2
5
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17
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27
7
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6
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51
6
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51
6
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6061
Gulzar
-
Preparation of N-substituted a ...
Escherichia coli
J. Chem. Soc. Perkin Trans. I
1197
649-655
1997
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6059
Kato
3-Methylaspartate ammonia-lyas ...
Citrobacter amalonaticus, Citrobacter amalonaticus YG-1002, Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920, Clostridium tetanomorphum NCIMB 11547
Appl. Microbiol. Biotechnol.
43
901-907
1995
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22
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2
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23
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31
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31
2
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2
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6060
Kato
Purification and properties of ...
Citrobacter freundii, Clostridium tetanomorphum, Clostridium tetanomorphum NCIMB 11547, Morganella morganii, Morganella morganii YG-0601
Biosci. Biotechnol. Biochem.
59
93-99
1995
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2
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33
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20
4
5
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34
3
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3
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33
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34
3
3
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8
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3
4
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6057
Goda
Cloning, sequencing, and expre ...
Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920
Biochemistry
31
10747-10756
1992
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1
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6058
Botting
Mechanism of C-3 hydrogen exch ...
Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920
Biochemistry
31
1509-1520
1992
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13
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6055
Botting
Substrate specificity of the 3 ...
Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920
Biochemistry
27
2953-2955
1988
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13
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6056
Botting
Primary deuterium isotope effe ...
Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920
Biochemistry
27
2956-2959
1988
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12
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6054
Akhtar
-
Enantiospecific synthesis of 3 ...
Clostridium tetanomorphum
Tetrahedron
43
5899-5908
1987
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8
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8
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6066
Botting
-
Mechanism of the enzymic elimi ...
Bacterium cadaveris, Clostridium tetanomorphum
J. Chem. Soc. Chem. Commun.
1987
1371-1373
1987
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2
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2
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11
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11
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6067
Akhtar
-
Stereochemical course of the e ...
Bacterium cadaveris, Clostridium tetanomorphum
Tetrahedron Lett.
28
2413-2416
1987
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2
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2
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11
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6053
Akhtar
-
Enzymic synthesis of 3-halogen ...
Clostridium tetanomorphum
J. Chem. Soc. Chem. Commun.
1986
1290-1291
1980
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1
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6
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6
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-
5997
Hanson
-
The enzymic elimination of amm ...
Bacterium cadaveris, Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
7
75-166
1972
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3
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9
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9
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6051
Hsiang
-
beta-methylaspartase from Clos ...
Bacterium cadaveris, Clostridium tetanomorphum, Clostridium tetanomorphum H1 / ATCC 15920
Methods Enzymol.
13
347-353
1969
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3
1
2
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13
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1
17
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2
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1
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1
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3
1
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1
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1
1
17
-
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2
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6050
Bright
Divalent metal activation of b ...
Clostridium tetanomorphum
Biochemistry
6
1191-1203
1967
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3
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9
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2
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6065
Winkler
New substrates for beta-methyl ...
Clostridium tetanomorphum
Biochim. Biophys. Acta
146
287-289
1967
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10
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-
6049
Bright
-
The preparation of crystalline ...
Clostridium tetanomorphum
Biochim. Biophys. Acta
44
586-588
1960
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5
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