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Literature summary for 1.1.1.376 extracted from

  • Watanabe, S.; Kodaki, T.; Kodak, T.; Makino, K.
    Cloning, expression, and characterization of bacterial L-arabinose 1-dehydrogenase involved in an alternative pathway of L-arabinose metabolism (2006), J. Biol. Chem., 281, 2612-2623.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in Escherichia coli Azospirillum brasilense

Protein Variants

Protein Variants Comment Organism
D168A mutant shows no activity under standard assay conditions Azospirillum brasilense
N172A kinetic analysis shows that the N172A mutant decreases by 4 orders of magnitude in kcat/Km values, compared with the wild-type enzyme Azospirillum brasilense

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.109
-
D-galactose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
0.255
-
L-arabinose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
0.26
-
L-arabinose pH 9.0, 30°C, cofactor: NADP+, recombinant wild-type enzyme Azospirillum brasilense
0.785
-
L-arabinose pH 9.0, 30°C, cofactor: NAD+, recombinant wild-type enzyme Azospirillum brasilense
1.41
-
L-arabinose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
1.49
-
D-galactose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
2 10 D-xylose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
3.95
-
D-talose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
5.87
-
D-talose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
28.9
-
L-arabinose pH 9.0, 30°C, cofactor: NADP+, mutant enzyme N172A Azospirillum brasilense
42.5
-
L-arabinose pH 9.0, 30°C, cofactor: NAD+, mutant enzyme N172A Azospirillum brasilense
72
-
D-xylose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense

Metals/Ions

Metals/Ions Comment Organism Structure
additional information no significant increase in activity in the presence of MgCl2, MnCl2, ZnCl2, CoCl2, NiCl2, or CaCl2 at final concentrations of 1 mM Azospirillum brasilense

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
33664
-
1 * 33664, calculated from sequence Azospirillum brasilense
39500
-
1 * 39500, SDS-PAGE Azospirillum brasilense
46400
-
gel filtration Azospirillum brasilense

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-arabinose + NADP+ Azospirillum brasilense first step of L-arabinose metabolism. The enzyme is involved in the metabolism of L-arabinose but not D-galactose L-arabinono-1,4-lactone + NADPH + H+
-
?

Organism

Organism UniProt Comment Textmining
Azospirillum brasilense Q53TZ2
-
-
Azospirillum brasilense DSM 1690 Q53TZ2
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Azospirillum brasilense

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.042
-
pH 9.0, 30°C, L-arabinose + NAD+, mutant enzyme N172A Azospirillum brasilense
0.148
-
pH 9.0, 30°C, L-arabinose + NADP+, mutant enzyme N172A Azospirillum brasilense
1.7
-
pH 9.0, 30°C, D-talose + NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
5.3
-
pH 9.0, 30°C, D-xylose + NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
12.8
-
pH 9.0, 30°C, D-talose + NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
14.8
-
pH 9.0, 30°C, D-xylose + NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
14.9
-
pH 9.0, 30°C, L-arabinose + NAD+, recombinant wild-type enzyme Azospirillum brasilense
23.8
-
pH 9.0, 30°C, D-galactose + NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
25
-
pH 9.0, 30°C, L-arabinose + NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
32
-
pH 9.0, 30°C, L-arabinose + NADP+, recombinant wild-type enzyme Azospirillum brasilense
35.6
-
pH 9.0, 30°C, D-galactose + NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
44.9
-
pH 9.0, 30°C, L-arabinose + NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-galactose + NAD+ the enzyme shows high catalytic efficiency for both L-arabinose and D-galactose. The enzyme prefers NADP+ over NAD+ Azospirillum brasilense ?
-
?
D-galactose + NAD+ the enzyme shows high catalytic efficiency for both L-arabinose and D-galactose. The enzyme prefers NADP+ over NAD+ Azospirillum brasilense DSM 1690 ?
-
?
D-galactose + NADP+ the enzyme shows high catalytic efficiency for both L-arabinose and D-galactose. The enzyme prefers NADP+ over NAD+ Azospirillum brasilense ?
-
?
D-galactose + NADP+ the enzyme shows high catalytic efficiency for both L-arabinose and D-galactose. The enzyme prefers NADP+ over NAD+ Azospirillum brasilense DSM 1690 ?
-
?
D-talose + NAD+ low activity Azospirillum brasilense ?
-
?
D-talose + NADP+ low activity Azospirillum brasilense ?
-
?
D-xylose + NAD+ low activity Azospirillum brasilense ?
-
?
D-xylose + NAD+ low activity Azospirillum brasilense DSM 1690 ?
-
?
D-xylose + NADP+ low activity Azospirillum brasilense ?
-
?
D-xylose + NADP+ low activity Azospirillum brasilense DSM 1690 ?
-
?
L-arabinose + NAD+ the enzyme shows high catalytic efficiency for both L-arabinose and D-galactose. The enzyme prefers NADP+ over NAD+ Azospirillum brasilense L-arabinono-1,4-lactone + NADH + H+
-
?
L-arabinose + NADP+ first step of L-arabinose metabolism. The enzyme is involved in the metabolism of L-arabinose but not D-galactose Azospirillum brasilense L-arabinono-1,4-lactone + NADPH + H+
-
?
L-arabinose + NADP+ the enzyme shows high catalytic efficiency for both L-arabinose and D-galactose. The enzyme prefers NADP+ over NAD+ Azospirillum brasilense L-arabinono-1,4-lactone + NADPH + H+
-
?
additional information no activity with D-arabinose, D-glucose, D-ribose, L-xylose, L-mannose, L-lyxose, and D-fructose (less than 1% of the activity with L-arabinose) Azospirillum brasilense ?
-
?
additional information no activity with D-arabinose, D-glucose, D-ribose, L-xylose, L-mannose, L-lyxose, and D-fructose (less than 1% of the activity with L-arabinose) Azospirillum brasilense DSM 1690 ?
-
?

Subunits

Subunits Comment Organism
monomer 1 * 39500, SDS-PAGE Azospirillum brasilense
monomer 1 * 33664, calculated from sequence Azospirillum brasilense

Synonyms

Synonyms Comment Organism
L-arabinose 1-dehydrogenase ambiguous Azospirillum brasilense

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Azospirillum brasilense

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.1
-
L-arabinose pH 9.0, 30°C, cofactor: NAD+, mutant enzyme N172A Azospirillum brasilense
0.3
-
L-arabinose pH 9.0, 30°C, cofactor: NADP+, mutant enzyme N172A Azospirillum brasilense
1.04
-
D-talose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
4
-
D-xylose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
6.1
-
L-arabinose pH 9.0, 30°C, cofactor: NAD+, recombinant wild-type enzyme Azospirillum brasilense
9.7
-
D-talose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
12
-
D-xylose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
16.5
-
L-arabinose pH 9.0, 30°C, cofactor: NADP+, recombinant wild-type enzyme Azospirillum brasilense
19
-
L-arabinose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
19.8
-
D-galactose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
26
-
D-galactose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
33.3
-
L-arabinose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
assay at Azospirillum brasilense

Cofactor

Cofactor Comment Organism Structure
NAD+ the enzyme prefers NADP+ over NAD+ Azospirillum brasilense
NADP+ the enzyme prefers NADP+ over NAD+ Azospirillum brasilense

Expression

Organism Comment Expression
Azospirillum brasilense the gene is induced by L-arabinose but not by D-galactose up

General Information

General Information Comment Organism
malfunction a disruptant of the L-arabinose 1-dehydrogenase gene does not grow on L-arabinose but grows on D-galactose at the same growth rate as the wild-type strain Azospirillum brasilense

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.01
-
L-arabinose pH 9.0, 30°C, cofactor: NADP+, mutant enzyme N172A Azospirillum brasilense
0.019
-
D-xylose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
0.03
-
L-arabinose pH 9.0, 30°C, cofactor: NAD+, mutant enzyme N172A Azospirillum brasilense
0.17
-
D-xylose pH 9.0, 30°C, cofactor: NADP*, enzyme purified from Azospirillum brasilense Azospirillum brasilense
0.26
-
D-talose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
1.6
-
D-talose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
7.78
-
L-arabinose pH 9.0, 30°C, cofactor: NAD+, recombinant wild-type enzyme Azospirillum brasilense
13.3
-
D-galactose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
13.4
-
L-arabinose pH 9.0, 30°C, cofactor: NAD+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
63.5
-
L-arabinose pH 9.0, 30°C, cofactor: NADP+, recombinant wild-type enzyme Azospirillum brasilense
131
-
L-arabinose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense
240
-
D-galactose pH 9.0, 30°C, cofactor: NADP+, enzyme purified from Azospirillum brasilense Azospirillum brasilense