Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.14.12.24 extracted from

  • Keenan, B.G.; Leungsakul, T.; Smets, B.F.; Wood, T.K.
    Saturation mutagenesis of Burkholderia cepacia R34 2,4-dinitrotoluene dioxygenase at DntAc valine 350 for synthesizing nitrohydroquinone, methylhydroquinone, and methoxyhydroquinone (2004), Appl. Environ. Microbiol., 70, 3222-3231.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.12.24

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli TG1 cells Burkholderia cepacia

Protein Variants

Protein Variants Comment Organism
V350F mutant displays significantly increased activity towards o-nitrophenol (47 times), m-nitrophenol (34 times), and o-methoxyphenol (174 times) as well as an expanded substrate range that now includes m-methoxyphenol, o-cresol, and m-cresol Burkholderia sp.
V350F the mutant of the alpha subunit is inactive with 2,4-dinitrotoluene but shows significantly increased activity towards 2-nitrophenol (47times), 3-nitrophenol (34times), and 2-methoxyphenol (174times) as well as an expanded substrate range that now includes 3-methoxyphenol, o-cresol, and m-cresol (wild type enzyme has no detectable activity for these substrates). The mutant also exhibits 10fold enhanced activity towards naphthalene forming (1R,2S)-cis-1,2-dihydro-1,2-dihydroxynaphthalene Burkholderia cepacia
V350M mutant displays increased activity towards o-nitrophenol (20 times) and o-methoxyphenol (162 times) as well as novel activity towards o-cresol Burkholderia sp.
V350M the mutant is inactive with 2,4-dinitrotoluene but displays increased activity towards 2-nitrophenol (20times) and 2-methoxyphenol (162times) as well as novel activity towards 2-cresol compared to the wild type enzyme. The mutant also exhibits 10fold enhanced activity towards naphthalene forming (1R,2S)-cis-1,2-dihydro-1,2-dihydroxynaphthalene Burkholderia cepacia

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2,4-dinitrotoluene + NADH + O2 Burkholderia cepacia highest activity 4-methyl-5-nitrocatechol + nitrite + NAD+
-
 ?
2,4-dinitrotoluene + NADH + O2 Burkholderia cepacia R34 highest activity 4-methyl-5-nitrocatechol + nitrite + NAD+
-
 ?

Organism

Organism UniProt Comment Textmining
Burkholderia cepacia
-
-
-
Burkholderia cepacia R34
-
-
-
Burkholderia sp. Q5BM20 and Q6QUY4 and Q45695 and Q6QUY3 Q5BM20 i.e flavoprotein reductase DntAa, Q6QUY4 i.e iron-sulfur ferredoxin DntAb, Q45695 i.e. large subunit DntAc, Q6QUY3 i.e. small subunit DntAd
-
Burkholderia sp. DNT Q5BM20 and Q6QUY4 and Q45695 and Q6QUY3 Q5BM20 i.e flavoprotein reductase DntAa, Q6QUY4 i.e iron-sulfur ferredoxin DntAb, Q45695 i.e. large subunit DntAc, Q6QUY3 i.e. small subunit DntAd
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,4-dinitrotoluene + NADH + H+ + O2
-
 Burkholderia sp. 4-methyl-5-nitrocatechol + nitrite + NAD+
-
 ?
2,4-dinitrotoluene + NADH + H+ + O2
-
 Burkholderia sp. DNT 4-methyl-5-nitrocatechol + nitrite + NAD+
-
 ?
2,4-dinitrotoluene + NADH + O2 highest activity Burkholderia cepacia 4-methyl-5-nitrocatechol + nitrite + NAD+
-
 ?
2,4-dinitrotoluene + NADH + O2 highest activity Burkholderia cepacia R34 4-methyl-5-nitrocatechol + nitrite + NAD+
-
 ?
2-methoxyphenol + NADH + O2
-
 Burkholderia cepacia methoxyhydroquinone + NAD+ + H2O
-
 ?
2-methoxyphenol + NADH + O2
-
 Burkholderia cepacia R34 methoxyhydroquinone + NAD+ + H2O
-
 ?
2-nitrophenol + NADH + O2
-
 Burkholderia cepacia nitrohydroquinone + 3-nitrocatechol + NAD+ + H2O
-
 ?
2-nitrophenol + NADH + O2
-
 Burkholderia cepacia R34 nitrohydroquinone + 3-nitrocatechol + NAD+ + H2O
-
 ?
3-nitrophenol + NADH + O2
-
 Burkholderia cepacia 4-nitrocatechol + NAD+ + H2O
-
 ?
3-nitrophenol + NADH + O2
-
 Burkholderia cepacia R34 4-nitrocatechol + NAD+ + H2O
-
 ?
m-cresol + NADH + H+ + O2
-
 Burkholderia sp. methylhydroquinone + nitrite + NAD+ no substrate for wild-type. Mutant V350F produces methylhydroquinone at 0.43 nmol/min/mg of protein ?
m-cresol + NADH + H+ + O2
-
 Burkholderia sp. DNT methylhydroquinone + nitrite + NAD+ no substrate for wild-type. Mutant V350F produces methylhydroquinone at 0.43 nmol/min/mg of protein ?
m-methoxyphenol + NADH + H+ + O2
-
 Burkholderia sp. methoxyhydroquinone + nitrite + NAD+ no substrate for wild-type. Mutant V350F produces methoxyhydroquinone at 0.55 nmol/min/mg of protein ?
m-methoxyphenol + NADH + H+ + O2
-
 Burkholderia sp. DNT methoxyhydroquinone + nitrite + NAD+ no substrate for wild-type. Mutant V350F produces methoxyhydroquinone at 0.55 nmol/min/mg of protein ?
m-nitrophenol + NADH + H+ + O2
-
 Burkholderia sp. 4-nitrocatechol + nitrite + NAD+ no substrate for wild-type. Mutant V350F produces 4-nitrocatechol at 0.29 nmol/min/mg of protein ?
m-nitrophenol + NADH + H+ + O2
-
 Burkholderia sp. DNT 4-nitrocatechol + nitrite + NAD+ no substrate for wild-type. Mutant V350F produces 4-nitrocatechol at 0.29 nmol/min/mg of protein ?
additional information no activity with 2-cresol, 3-methoxyphenol Burkholderia cepacia ?
-
 ?
additional information no activity with 2-cresol, 3-methoxyphenol Burkholderia cepacia R34 ?
-
 ?
naphthalene + NADH + H+ + O2
-
 Burkholderia sp. (1R,2S)-cis-1,2-dihydroxy-1,2-dihydronaphthalene + NAD+ mutants V350F and V350M exhibit 10fold enhanced activity towards naphthalene ?
naphthalene + NADH + H+ + O2
-
 Burkholderia sp. DNT (1R,2S)-cis-1,2-dihydroxy-1,2-dihydronaphthalene + NAD+ mutants V350F and V350M exhibit 10fold enhanced activity towards naphthalene ?
naphthalene + NADH + O2
-
 Burkholderia cepacia (1R,3S)-cis-1,2-dihydro-1,2-dihydroxynaphthalene + NAD+ + H2O
-
 ?
o-cresol + NADH + H+ + O2
-
 Burkholderia sp. methylhydroquinone + 2-hydroxybenzyl alcohol + nitrite + NAD+ mutant V350F produces methylhydroquinone at 1.52 nmol/min/mg of protein and 2-hydroxybenzyl alcohol at 0.74 nmol/min/mg of protein. Mutant V350M produces methylhydroquinone at 1.97 nmol/min/mg of protein and 2-hydroxybenzyl alcohol at 0.11 nmol/min/mg of protein ?
o-methoxyphenol + NADH + H+ + O2
-
 Burkholderia sp. methoxyhydroquinone + nitrite + NAD+ mutant V350F produces methoxyhydroquinone at 2.5 nmol/min/mg of protein. Mutant V350M produces methoxyhydroquinone at 2.4 mol/min/mg of protein ?
o-nitrophenol + NADH + H+ + O2
-
 Burkholderia sp. nitrohydroquinone + 3-nitrocatechol + nitrite + NAD+ product of wild-type. Mutant V350F produces both nitrohydroquinone at a rate of 0.75 nmol/min/mg of protein and 3-nitrocatechol at a rate of 0.069 nmol/min/mg of protein from o-nitrophenol. Mutant V350M produces both nitrohydroquinone at 0.33 nmol/min/mg of protein and 3-nitrocatechol at 0.089 nmol/min/mg of protein ?

Synonyms

Synonyms Comment Organism
DDO
-
 Burkholderia cepacia
DntAc alpha subunit Burkholderia cepacia