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Literature summary for 1.1.1.203 extracted from

  • Roth, T.; Beer, B.; Pick, A.; Sieber, V.
    Thermostabilization of the uronate dehydrogenase from Agrobacterium tumefaciens by semi-rational design (2017), AMB Express, 7, 103 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.1.203

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
A41P/H101N/H236K the mutant shows 52.3% catalytic efficiency as compared to the wild type enzyme Agrobacterium tumefaciens
A41P/H101N/H236R the mutant shows 59.3% catalytic efficiency as compared to the wild type enzyme Agrobacterium tumefaciens
A41P/H101Y/H236K the mutant shows 58.4% catalytic efficiency, higher kinetic and thermodynamic stability with a T50 value of 62.2°C after 15 min (3.2°C improvement) as compared to the wild type enzyme Agrobacterium tumefaciens
A41P/H101Y/H236R the mutant shows 56% catalytic efficiency as compared to the wild type enzyme Agrobacterium tumefaciens
A41P/H236K the mutant shows 62.7% catalytic efficiency as compared to the wild type enzyme Agrobacterium tumefaciens
H101N/H236K the mutant shows 48.8% catalytic efficiency as compared to the wild type enzyme Agrobacterium tumefaciens
H101Y/H236K the mutant shows 40.8% catalytic efficiency as compared to the wild type enzyme Agrobacterium tumefaciens
H236K the mutant shows 34% catalytic efficiency as compared to the wild type enzyme Agrobacterium tumefaciens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.7
-
 beta-D-glucuronate wild type enzyme, at pH 8.0 and 25°C Agrobacterium tumefaciens
0.9
-
 beta-D-glucuronate mutant enzyme A41P/H101N/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
0.9
-
 beta-D-glucuronate mutant enzyme A41P/H101Y/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
0.9
-
 beta-D-glucuronate mutant enzyme H101N/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
1
-
 beta-D-glucuronate mutant enzyme A41P/H101N/H236R, at pH 8.0 and 25°C Agrobacterium tumefaciens
1
-
 beta-D-glucuronate mutant enzyme A41P/H101Y/H236R, at pH 8.0 and 25°C Agrobacterium tumefaciens
1
-
 beta-D-glucuronate mutant enzyme H101Y/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
1.2
-
 beta-D-glucuronate mutant enzyme A41P/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
1.3
-
 beta-D-glucuronate mutant enzyme H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ 5 mM used in assay conditions Agrobacterium tumefaciens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
beta-D-galacturonate + NAD+ Agrobacterium tumefaciens
-
 D-galactaro-1,5-lactone + NADH + H+
-
 ?
beta-D-glucuronate + NAD+ Agrobacterium tumefaciens
-
 D-glucaro-1,5-lactone + NADH + H+
-
 ?

Organism

Organism UniProt Comment Textmining
Agrobacterium tumefaciens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap column chromatography Agrobacterium tumefaciens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
218
-
 mutant enzyme H101Y/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
228
-
 mutant enzyme H101N/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
234
-
 mutant enzyme H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
248
-
 mutant enzyme A41P/H101N/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
280
-
 mutant enzyme A41P/H101Y/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
302
-
 mutant enzyme A41P/H101Y/H236R, at pH 8.0 and 25°C Agrobacterium tumefaciens
330
-
 mutant enzyme A41P/H101N/H236R, at pH 8.0 and 25°C Agrobacterium tumefaciens
391
-
 wild type enzyme, at pH 8.0 and 25°C Agrobacterium tumefaciens
407
-
 mutant enzyme A41P/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
beta-D-galacturonate + NAD+
-
 Agrobacterium tumefaciens D-galactaro-1,5-lactone + NADH + H+
-
 ?
beta-D-glucuronate + NAD+
-
 Agrobacterium tumefaciens D-glucaro-1,5-lactone + NADH + H+
-
 ?

Subunits

Subunits Comment Organism
? x * 31210, calculated from amino acid sequence Agrobacterium tumefaciens

Synonyms

Synonyms Comment Organism
Udh
-
 Agrobacterium tumefaciens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
59
-
 the enzyme activity is reduced to 50% after 15 min at 59°C Agrobacterium tumefaciens

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Agrobacterium tumefaciens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
97.5
-
 beta-D-glucuronate mutant enzyme H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
116.7
-
 beta-D-glucuronate mutant enzyme H101Y/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
139.7
-
 beta-D-glucuronate mutant enzyme H101N/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
149.7
-
 beta-D-glucuronate mutant enzyme A41P/H101N/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
160.5
-
 beta-D-glucuronate mutant enzyme A41P/H101Y/H236R, at pH 8.0 and 25°C Agrobacterium tumefaciens
167.3
-
 beta-D-glucuronate mutant enzyme A41P/H101Y/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
170
-
 beta-D-glucuronate mutant enzyme A41P/H101N/H236R, at pH 8.0 and 25°C Agrobacterium tumefaciens
179.5
-
 beta-D-glucuronate mutant enzyme A41P/H236K, at pH 8.0 and 25°C Agrobacterium tumefaciens
286.4
-
 beta-D-glucuronate wild type enzyme, at pH 8.0 and 25°C Agrobacterium tumefaciens