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Literature summary for 1.7.1.6 extracted from

  • Lu, L.; Yang, Y.; Lin, H.; Gao, F.; Zhao, Y.
    Construction, expression and characterization of fusion enzyme containing azoreductase and glucose 1-dehydrogenase for dye removal (2014), Int. Biodeter. Biodegrad., 87, 81-86 .
No PubMed abstract available
Search for more literature for 1.7.1.6

Activating Compound

Activating Compound Comment Organism Structure
dodecyl trimethyl ammonium bromide
-
 Shewanella oneidensis
sodium dodecyl benzene sulfonate
-
 Shewanella oneidensis

Cloned(Commentary)

Cloned (Comment) Organism
functional recombinant expression of the chimeric gene GFA, in which both GDH stop codon and the sequence encoding AZR signal peptide are removed, in Escherichia coli strain BL21 from plasmid pET28a(+) Shewanella oneidensis

Protein Variants

Protein Variants Comment Organism
additional information construction of a chimeric artificial bifunctional enzyme chimeric enzyme composed of the two oligomeric enzymes, azoreductase (AZR) from Shewanella oneidensis MR-1 and the alkaline-resistant NAD(P)-dependent glucose 1-dehydrogenase (LsGDH) from Lysinibacillus sphaericus G10. The reaction is extended to a recycling system. The fusion enzyme exhibits a higher stability and suitability for high-temperature application than the parental wild-type enzyme Shewanella oneidensis

Inhibitors

Inhibitors Comment Organism Structure
Cd2+ complete inhibition at 25 mM Shewanella oneidensis
cetyl dimethylethyl ammonium bromide
-
 Shewanella oneidensis
Cetylpyridinium chloride
-
 Shewanella oneidensis
Mn2+ inhibits at 25 mM Shewanella oneidensis
SDS
-
 Shewanella oneidensis
tetradecylammonium bromide
-
 Shewanella oneidensis
Zn2+ inhibits at 25 mM Shewanella oneidensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00476
-
 Methyl red recombinant chimeric fusion protein AZR-GDH, pH 7.0, 40°C Shewanella oneidensis
0.00916
-
 Methyl red recombinant wild-type AZR, pH 7.0, 40°C Shewanella oneidensis

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ activates 1.84fold at 25 mM Shewanella oneidensis
Mg2+ activates 1.28fold at 25 mM Shewanella oneidensis

Organism

Organism UniProt Comment Textmining
Shewanella oneidensis Q8EBE0
-
-
Shewanella oneidensis MR-1 / ATCC 700550 Q8EBE0
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant chimeric fusion protein AZR-GDH from Escherichia coli strain BL21 Shewanella oneidensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
153.16
-
 purified wild-type AZR, 40°C, pH 7.0, substrate methyl red Shewanella oneidensis
160.62
-
 purified recombinant fusion protein AZR-GDH, 40°C, pH 7.0, substrate methyl red Shewanella oneidensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
methyl red + 2 NADH + 2 H+ i.e. 4-dimethylaminoazobenzene-2'-carboxylic acid Shewanella oneidensis anthranilate + 4-amino-N,N-dimethylaniline + 2 NAD+
-
 ?
methyl red + 2 NADH + 2 H+ i.e. 4-dimethylaminoazobenzene-2'-carboxylic acid Shewanella oneidensis MR-1 / ATCC 700550 anthranilate + 4-amino-N,N-dimethylaniline + 2 NAD+
-
 ?

Subunits

Subunits Comment Organism
? x * 55000, recombinant chimeric fusion protein AZR-GDH, SDS-PAGE Shewanella oneidensis

Synonyms

Synonyms Comment Organism
azoreductase
-
 Shewanella oneidensis
AZR
-
 Shewanella oneidensis
NADPH-dependent azoreductase Azr UniProt Shewanella oneidensis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
 recombinant chimeric fusion protein AZR-GDH Shewanella oneidensis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 60 recombinant chimeric fusion protein AZR-GDH, activity range, 63.2% and 4.0% of maximal activity at 55°C and 60°C, respectively. The residual activity of the parental wild-type azoreductase decreases to 1% at 55°C Shewanella oneidensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
82.17
-
 Methyl red recombinant wild-type AZR, pH 7.0, 40°C Shewanella oneidensis
102.92
-
 Methyl red recombinant chimeric fusion protein AZR-GDH, pH 7.0, 40°C Shewanella oneidensis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 recombinant chimeric fusion protein AZR-GDH Shewanella oneidensis

pH Range

pH Minimum pH Maximum Comment Organism
4 9 recombinant chimeric fusion protein AZR-GDH, activity range, profile overview Shewanella oneidensis

pH Stability

pH Stability pH Stability Maximum Comment Organism
4 8 completely stable at pH 7.0-7.5, 50% activity remaining at pH 5.0 and pH 7.8, 20% activity remaining at pH 4.0 and pH 8.0 Shewanella oneidensis
7 7.5 purified recombinant chimeric fusion protein AZR-GDH, completely stable at Shewanella oneidensis

Cofactor

Cofactor Comment Organism Structure
NADH
-
 Shewanella oneidensis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
8970.5
-
 Methyl red recombinant wild-type AZR, pH 7.0, 40°C Shewanella oneidensis
21621.8
-
 Methyl red recombinant chimeric fusion protein AZR-GDH, pH 7.0, 40°C Shewanella oneidensis