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Literature summary for 1.5.1.37 extracted from

  • Gisi, M.R.; Xun, L.
    Characterization of chlorophenol 4-monooxygenase (TftD) and NADH:flavin adenine dinucleotide oxidoreductase (TftC) of Burkholderia cepacia AC1100 (2003), J. Bacteriol., 185, 2786-2792.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli as a C-terminally His-tagged fusion protein Paraburkholderia phenoliruptrix

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0048
-
FAD cosubstrate: NADH, pH 7.0, 24°C Paraburkholderia phenoliruptrix
0.0103
-
FMN cosubstrate: NADH, pH 7.0, 24°C Paraburkholderia phenoliruptrix
0.0401
-
NADH cosubstrate: FAD, pH 7.0, 24°C Paraburkholderia phenoliruptrix
0.164
-
NADH cosubstrate: FMN, pH 7.0, 24°C Paraburkholderia phenoliruptrix

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
22000
-
x * 22000, His-tagged TftC, SDS-PAGE Paraburkholderia phenoliruptrix

Organism

Organism UniProt Comment Textmining
Paraburkholderia phenoliruptrix
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Paraburkholderia phenoliruptrix

Storage Stability

Storage Stability Organism
-80°C, His-tagged TftC loses about 50% activity after 2 months Paraburkholderia phenoliruptrix
0°C, His-tagged TftC is unstable in the elution buffer containing 140 mM imidazole on ice, losing about 50% activity in 2 h Paraburkholderia phenoliruptrix

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
FAD + NADH + H+ FAD is a better substrate than FMN. The enzyme does not use NADPH or riboflavin as substrate Paraburkholderia phenoliruptrix FADH2 + NAD+
-
?
FMN + NADH + H+ FAD is a better substrate than FMN. The enzyme does not use NADPH or riboflavin as substrate Paraburkholderia phenoliruptrix FMNH2 + NAD+
-
?

Subunits

Subunits Comment Organism
? x * 22000, His-tagged TftC, SDS-PAGE Paraburkholderia phenoliruptrix

Synonyms

Synonyms Comment Organism
NADH-dependent FAD reductase
-
Paraburkholderia phenoliruptrix
NADH-FAD reductase
-
Paraburkholderia phenoliruptrix
NADH:FAD oxidoreductase
-
Paraburkholderia phenoliruptrix
NADH:flavin adenine dinucleotide oxidoreductase
-
Paraburkholderia phenoliruptrix
TftC
-
Paraburkholderia phenoliruptrix

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
24
-
assay at Paraburkholderia phenoliruptrix
30
-
-
Paraburkholderia phenoliruptrix

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
23 45 23°C: 85% of maximal activity, 45°C: 70% of maximal activity Paraburkholderia phenoliruptrix

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
11.9
-
NADH cosubstrate: FMN, pH 7.0, 24°C Paraburkholderia phenoliruptrix
16.6
-
FAD cosubstrate: NADH, pH 7.0, 24°C Paraburkholderia phenoliruptrix
18.5
-
FMN cosubstrate: NADH, pH 7.0, 24°C Paraburkholderia phenoliruptrix
30.1
-
NADH cosubstrate: FAD, pH 7.0, 24°C Paraburkholderia phenoliruptrix

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
Na-phosphate buffer Paraburkholderia phenoliruptrix
7
-
assay at Paraburkholderia phenoliruptrix

pH Range

pH Minimum pH Maximum Comment Organism
5 7.6 pH 5.0 (sodium succinate buffer): 52% of maximal activity, pH 7.6: 75% of maximal activity Paraburkholderia phenoliruptrix

Cofactor

Cofactor Comment Organism Structure
NADH the enzyme does not use NADPH as substrate Paraburkholderia phenoliruptrix

General Information

General Information Comment Organism
physiological function TftD supplies FADH2 to (FADH2)-utilizing monooxygenase TftD, that converts 2,4,5-trichlorophenol to 2,5-dichloro-p-quinol and then to 5-chlorohydroxyquinol. It also converts 2,4,6-trichlorophenol to 2,6-dichloro-p-quinol as the final product Paraburkholderia phenoliruptrix

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
72.5
-
NADH cosubstrate: FMN, pH 7.0, 24°C Paraburkholderia phenoliruptrix
77.3
-
NADH cosubstrate: FAD, pH 7.0, 24°C Paraburkholderia phenoliruptrix
1796
-
FMN cosubstrate: NADH, pH 7.0, 24°C Paraburkholderia phenoliruptrix
3458
-
FAD cosubstrate: NADH, pH 7.0, 24°C Paraburkholderia phenoliruptrix