Information on EC 1.2.1.95 - L-2-aminoadipate reductase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.2.1.95
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RECOMMENDED NAME
GeneOntology No.
L-2-aminoadipate reductase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(S)-2-amino-6-oxohexanoate + holo-[LYS2 peptidyl-carrier-protein] + NADP+ = L-2-aminoadipyl-[LYS2 peptidyl-carrier-protein] + NADPH + H+
show the reaction diagram
(1b)
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-
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(S)-2-amino-6-oxohexanoate + NADP+ + AMP + diphosphate = L-2-aminoadipate + NADPH + H+ + ATP
show the reaction diagram
overall reaction
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-
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L-2-aminoadipyl-[LYS2 peptidyl-carrier-protein] + AMP + diphosphate = L-2-aminoadipate + holo-[LYS2 peptidyl-carrier-protein] + ATP
show the reaction diagram
(1a)
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Lysine biosynthesis
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
(S)-2-amino-6-oxohexanoate:NADP+ oxidoreductase (ATP-forming)
This enzyme, characterized from the yeast Saccharomyces cerevisiae, catalyses the reduction of L-2-aminoadipate to (S)-2-amino-6-oxohexanoate during L-lysine biosynthesis. An adenylation domain activates the substrate at the expense of ATP hydrolysis, and forms L-2-aminoadipate adenylate, which is attached to a peptidyl-carrier protein (PCP) domain. Binding of NADPH results in reductive cleavage of the acyl-S-enzyme intermediate, releasing (S)-2-amino-6-oxohexanoate. Different from EC 1.2.1.31, L-aminoadipate-semialdehyde dehydrogenase, which catalyses a similar transformation in the opposite direction without ATP hydrolysis.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
the enzyme is involved in the fungal de novo L-lysine biosynthesis via ATP- and NADPH-dependent reduction of the intermediate L-alpha-aminoadipic acid into L-alpha-aminoadipate 6-semialdehyde as a multifunctional aminoacyl-adenylate-forming reductase, pathway overview
physiological function
L-alpha-aminoadipic acid reductases catalyze the ATP- and NADPH-dependent reduction of L-alpha-aminoadipic acid to the corresponding 6-semialdehyde during fungal L-lysine biosynthesis
additional information
the enzyme has a multidomain composition but features a unique domain of elusive function, termed the adenylation activating (ADA) domain, that extends the reductase N-terminally. The activity of alpha-aminoadipate reductase and A domain depends on the N-terminally extending domain. ADA domain sequence comparison and protein interaction analysis, homology modeling, overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-2-amino-6-oxohexanoate + NADP+ + AMP + diphosphate
L-2-aminoadipate + NADPH + H+ + ATP
show the reaction diagram
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presence of ATP and Mg2+ required
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?
adipate + NADPH + H+ + ATP
adipate semialdehyde + NADP+ + AMP + diphosphate
show the reaction diagram
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-
-
?
D-2-aminoadipate + NADPH + H+ + ATP
(R)-2-amino-6-oxohexanoate + NADP+ + AMP + diphosphate
show the reaction diagram
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-
-
?
L-2-aminoadipate + NADPH + H+ + ATP
(S)-2-amino-6-oxohexanoate + NADP+ + AMP + diphosphate
show the reaction diagram
S-carboxymethyl-L-cysteine + NADPH + H+ + ATP
? + NADP+ + AMP + diphosphate
show the reaction diagram
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20fold lower activity than with L-2-aminoadipate
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-
?
S-carboxymethyl-L-cysteine + NADPH + H+ + ATP
S-formylmethyl-L-cysteine + NADP+ + AMP + diphosphate
show the reaction diagram
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-
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?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
L-2-aminoadipate + NADPH + H+ + ATP
(S)-2-amino-6-oxohexanoate + NADP+ + AMP + diphosphate
show the reaction diagram
A0A0S2LUS1
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?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
additional information
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
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absolutely required for activity, optimal concentration 10 mM
MgCl2
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required
Mn2+
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can partly replace Mg2+, leading to 60% of maximum activity with Mg2+
additional information
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no activity with Ca2+, Co2+, Hg2+, Cu2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
alpha-aminoadipate
inhibitory effect can be reversed by adding methionine
EDTA
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5 mM, 75% inhibition
L-lysine
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
10
adipate
pH 8.8, 37C
0.65 - 1.3
ATP
2.7
D-2-aminoadipate
pH 8.8, 37C
0.3 - 1.4
L-2-aminoadipate
0.06 - 0.62
NADPH
0.43
S-carboxymethyl-L-cysteine
pH 8.8, 37C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0045
adipate
pH 8.8, 37C
0.63
D-2-aminoadipate
pH 8.8, 37C
1.1 - 41.7
L-2-aminoadipate
6
S-carboxymethyl-L-cysteine
pH 8.8, 37C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0005
adipate
pH 8.8, 37C
0.23
D-2-aminoadipate
pH 8.8, 37C
88.3
L-2-aminoadipate
pH 8.8, 37C
14
S-carboxymethyl-L-cysteine
pH 8.8, 37C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.26
L-lysine
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pH not specified in the publication, temperature not specified in the publication
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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rapid loss of activity above
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
132600
about, recombinant truncated enzyme, sequence calculation
150000
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x * 150000, SDS-PAGE
154800
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1 * 155000, SDS-PAGE, 1 * 154800, calculated
155000
171000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
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1 * 155000, SDS-PAGE, 1 * 154800, calculated
additional information
the enzyme has a multidomain composition but features a unique domain of elusive function, termed the adenylation activating (ADA) domain, that extends the reductase N-terminally. The activity of alpha-aminoadipate reductase depends on the N-terminally extending domain
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
side-chain modification
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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1h, 20% loss of activity
37
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15 min, 50% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged truncated enzyme mutants from Escherichia coli strain KRX by nickel affinity chromatography and gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in ERscherichia coli
expression in Escherichia coli
expression of the ORF of the Saccharomycopsis fibuligera alpha-aminoadipate reductase gene (LYS2) in a lys2- strain of Saccharomyces cerevisiae
mutant enzymes expressed in Escherichia coli BL21
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recombinant expression of N-terminally His6-tagged truncated NPS3 enzyme mutant lacking the 247 amino acids of the N-terminal ADA domain from pET21a-based expression plasmid pMR1, recombinant expression of N-terminally His6-tagged truncated NPS3 enzyme mutant lacking the 152 amino acids of the N-terminal ADA domain from expression plasmid pDK22, and recombinant expression of the 102.8 kDa ADA-A didomain (lacking the C-terminal 515 amino acids) and the 75.7 kDa standalone NPS3A domain, respectively, in which the first 247 and terminal 515 amino acids are absent, from plasmids pDK24 and pDK25, both based on expression vector pET28a, all in Escherichia coli strain KRX
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
activity from cells grown in lysine-supplemented minimal or YEPD media is less than the activity of cells grown in minimal medium
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A459G
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106% of the alpha-aminoadipate reductase activity of wild-type enzyme
A475G
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109% of the alpha-aminoadipate reductase activity of wild-type enzyme
A979G
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94% of the alpha-aminoadipate reductase activity of wild-type enzyme
D461E
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4% of the alpha-aminoadipate reductase activity of wild-type enzyme
D461N
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no alpha-aminoadipate reductase activity
D466E
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15% of the alpha-aminoadipate reductase activity of wild-type enzyme
E420T
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101% of the alpha-aminoadipate reductase activity of wild-type enzyme
F415Y
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71% of the alpha-aminoadipate reductase activity of wild-type enzyme
F452L
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6% of the alpha-aminoadipate reductase activity of wild-type enzyme
F452V
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45% of the alpha-aminoadipate reductase activity of wild-type enzyme
F468L
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96% of the alpha-aminoadipate reductase activity of wild-type enzyme
F982L
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4% of the alpha-aminoadipate reductase activity of wild-type enzyme
F982V
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no alpha-aminoadipate reductase activity
G418A
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31% of the alpha-aminoadipate reductase activity of wild-type enzyme
G418V
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no alpha-aminoadipate reductase activity
G421A
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75% of the alpha-aminoadipate reductase activity of wild-type enzyme
G425A
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99% of the alpha-aminoadipate reductase activity of wild-type enzyme
G474A
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41% of the alpha-aminoadipate reductase activity of wild-type enzyme
G881E
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conserved residue in the posttranslational activation domain LGGHSI, less than 50% of wild-type activity
G882R
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conserved residue in the posttranslational activation domain LGGHSI, loss of activity
G882S
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conserved residue in the posttranslational activation domain LGGHSI, loss of activity
G978A
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2% of the alpha-aminoadipate reductase activity of wild-type enzyme
G978S
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no alpha-aminoadipate reductase activity
G981A
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no alpha-aminoadipate reductase activity
G981V
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no alpha-aminoadipate reductase activity
G984A
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3% of the alpha-aminoadipate reductase activity of wild-type enzyme
G984S
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no alpha-aminoadipate reductase activity
H460R
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no alpha-aminoadipate reductase activity
H883R
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conserved residue in the posttranslational activation domain LGGHSI, strong reduction of activity
I422V
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92% of the alpha-aminoadipate reductase activity of wild-type enzyme
I463L
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no alpha-aminoadipate reductase activity
I885L G881E
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conserved residue in the posttranslational activation domain LGGHSI, less than 50% of wild-type activity
I987V
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92% of the alpha-aminoadipate reductase activity of wild-type enzyme
K424R
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26% of the alpha-aminoadipate reductase activity of wild-type enzyme
K424T
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3% of the alpha-aminoadipate reductase activity of wild-type enzyme
K451Q
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58% of the alpha-aminoadipate reductase activity of wild-type enzyme
K451R
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no alpha-aminoadipate reductase activity
K535Q
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94% of the alpha-aminoadipate reductase activity of wild-type enzyme
L446I
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71% of the alpha-aminoadipate reductase activity of wild-type enzyme
L471V
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89% of the alpha-aminoadipate reductase activity of wild-type enzyme
L983V
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31% of the alpha-aminoadipate reductase activity of wild-type enzyme
M440L
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99% of the alpha-aminoadipate reductase activity of wild-type enzyme
P423V
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51% of the alpha-aminoadipate reductase activity of wild-type enzyme
P462A
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2% of the alpha-aminoadipate reductase activity of wild-type enzyme
P462S
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no alpha-aminoadipate reductase activity
Q464E
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17% of the alpha-aminoadipate reductase activity of wild-type enzyme
Q511E
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31% of the alpha-aminoadipate reductase activity of wild-type enzyme
Q511L
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129% of the alpha-aminoadipate reductase activity of wild-type enzyme
R465K
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2% of the alpha-aminoadipate reductase activity of wild-type enzyme
R465S
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no alpha-aminoadipate reductase activity
S417A
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10% of the alpha-aminoadipate reductase activity of wild-type enzyme
S417T
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82% of the alpha-aminoadipate reductase activity of wild-type enzyme
S419A
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16% of the alpha-aminoadipate reductase activity of wild-type enzyme
S419T
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no alpha-aminoadipate reductase activity
S456T
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59% of the alpha-aminoadipate reductase activity of wild-type enzyme
S884A
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conserved residue in the posttranslational activation domain LGGHSI, loss of activity
S884F
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conserved residue in the posttranslational activation domain LGGHSI, loss of activity
S985A
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85% of the alpha-aminoadipate reductase activity of wild-type enzyme
T416A
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no alpha-aminoadipate reductase activity
T416S
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35% of the alpha-aminoadipate reductase activity of wild-type enzyme
T469S
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80% of the alpha-aminoadipate reductase activity of wild-type enzyme
T506A
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49% of the alpha-aminoadipate reductase activity of wild-type enzyme
T506S
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no alpha-aminoadipate reductase activity
T534A
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103% of the alpha-aminoadipate reductase activity of wild-type enzyme
T977S
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70% of the alpha-aminoadipate reductase activity of wild-type enzyme
T980S
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28% of the alpha-aminoadipate reductase activity of wild-type enzyme
V426L
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64% of the alpha-aminoadipate reductase activity of wild-type enzyme
V529I
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117% of the alpha-aminoadipate reductase activity of wild-type enzyme
G910A
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mutation in the activation domain (IGGHSI), no activity
S913A
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mutation in the activation domain (IGGHSI), no activity
S913T
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mutation in the activation domain (IGGHSI), no activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information