1.14.13.236: toluene 4-monooxygenase

This is an abbreviated version!
For detailed information about toluene 4-monooxygenase, go to the full flat file.

Word Map on EC 1.14.13.236

Reaction

Toluene
+
NADH
+
H+
+
O2
=
4-Methylphenol
+
NAD+
+
H2O

Synonyms

T4moD, T4moF, T4MOH, TMO, TmoA, TmoC, TmoF, toluene-4-monooxygenase system protein A, TOM, TomA3

ECTree

     1 Oxidoreductases
         1.14 Acting on paired donors, with incorporation or reduction of molecular oxygen
             1.14.13 With NADH or NADPH as one donor, and incorporation of one atom of oxygen into the other donor
                1.14.13.236 toluene 4-monooxygenase

Engineering

Engineering on EC 1.14.13.236 - toluene 4-monooxygenase

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
V106A
-
initial thioanisole sulfoxidation is improved by 1.65fold
V106E
-
initial thioanisole sulfoxidation is improved by 1.72fold
V106L
-
initial thioanisole sulfoxidation is decreased by 0.43fold
V106M
-
mutant oxidizes methyl phenyl sulfide to the corresponding sulfoxide at a rate of 3.0 nmol/min/mg protein compared with 1.6 for the wild-type enzyme, and the enantiomeric excess (pro-S) increases from 51% for the wild type to 88% for this mutant. Function of residue V106 is the proper positioning or docking of the substrate with respect to the diiron atoms
V106S
-
initial thioanisole sulfoxidation is decreased by 0.8fold
V106A
-
initial thioanisole sulfoxidation is improved by 1.65fold
-
V106E
-
initial thioanisole sulfoxidation is improved by 1.72fold
-
V106L
-
initial thioanisole sulfoxidation is decreased by 0.43fold
-
V106M
-
mutant oxidizes methyl phenyl sulfide to the corresponding sulfoxide at a rate of 3.0 nmol/min/mg protein compared with 1.6 for the wild-type enzyme, and the enantiomeric excess (pro-S) increases from 51% for the wild type to 88% for this mutant. Function of residue V106 is the proper positioning or docking of the substrate with respect to the diiron atoms
-
V106S
-
initial thioanisole sulfoxidation is decreased by 0.8fold
-
D285A
-
mutation in subunit TmoA, 2.7fold increase in activity with 2-phenylethanol
D285C
-
mutation in subunit TmoA, 4fold increase in activity with 2-phenylethanol
D285I
-
mutation in subunit TmoA, 6.6fold increase in activity with 2-phenylethanol
D285L
-
mutation in subunit TmoA, 5.4fold increase in activity with 2-phenylethanol
D285P
-
mutation in subunit TmoA, 3.3fold increase in activity with 2-phenylethanol
D285Q
-
mutation in subunit TmoA, 10.5fold increase in activity with 2-phenylethanol
D285S
-
mutation in subunit TmoA, 70% of wild-type activity
D285Y mutation in subunit TmoA,
-
3fold increase in activity with 2-phenylethanol
F205I
decrease in regiospecificity for p-cresol formation, about 5-fold increase in the percentage of m-cresol formation. Mutant gives nearly equivalent amounts of benzylic and phenolic products from p-xylene oxidation
G103A/A107S
-
mutation in subunit TmoA, produces 3-methylcatechol (98%) from o-cresol twofold faster and produces 3-methoxycatechol (82%) from 1mM o-methoxyphenol seven times faster than the wild-type
G103S
-
mutation in subunit TmoA, produces 40fold more methoxyhydroquinone from o-methoxyphenol than the wild-type
G103S/A107T
-
mutation in subunit TmoA, produces methylhydroquinone (92%) from o-cresol fourfold faster than wild-type
I100A
I100A/D285I
-
mutation in subunit TmoA, 52fold increase in activity with 2-phenylethanol
I100A/D285Q
-
mutation in subunit TmoA, 85fold increase in activity with 2-phenylethanol
I100D
I100G
I100G/D285I
-
mutation in subunit TmoA, 14.1fold increase in activity with methyl p-tolyl sulfide
I100L
I100L/D285S
-
mutation in subunit TmoA, 1.4fold increase in activity with styrene
I100S
I100V
Q141C
decrease in regiospecificity for p-cresol formation, mutant functions predominantly as an aromatic ring hydroxylase during the oxidation of p-xylene
S395C
mutation in subunit TmoA, shows a 15fold increase in 2-phenylethanol hydroxylation rate
T201A
T201F
-
mutation causes a substantial shift in the product distribution, and gives o- and p-cresol in a 1:1 ratio
T201G
T201K
-
decrease in activity
T201L
parameters similar to wild-type
T201S
D285A
-
mutation in subunit TmoA, 2.7fold increase in activity with 2-phenylethanol
-
D285P
-
mutation in subunit TmoA, 3.3fold increase in activity with 2-phenylethanol
-
F205I
-
decrease in regiospecificity for p-cresol formation, about 5-fold increase in the percentage of m-cresol formation. Mutant gives nearly equivalent amounts of benzylic and phenolic products from p-xylene oxidation
-
G103A/A107S
-
mutation in subunit TmoA, produces 3-methylcatechol (98%) from o-cresol twofold faster and produces 3-methoxycatechol (82%) from 1mM o-methoxyphenol seven times faster than the wild-type
-
G103S
-
mutation in subunit TmoA, produces 40fold more methoxyhydroquinone from o-methoxyphenol than the wild-type
-
G103S/A107T
-
mutation in subunit TmoA, produces methylhydroquinone (92%) from o-cresol fourfold faster than wild-type
-
I100A
-
mutant hydroxylates o-tyrosol, m-tyrosol and p-tyrosol to form hydroxytyrosol; mutant shows similar rates as wild-type; mutation in subunit TmoA, 35fold increase in activity with 2-phenylethanol
-
I100D
-
mutation improves both reaction rate and enantioselectivity
-
I100G
-
mutant hydroxylates o-tyrosol, m-tyrosol and p-tyrosol to form hydroxytyrosol; mutation increases the wild-type oxidation rate of methyl phenyl sulfide by 1.7fold, and the enantiomeric excess rises from 86% to 98% pro-S. I100G oxidizes methyl para-tolyl sulfide 11 times faster than the wild type does and changes the selectivity from 41% pro-R to 77% pro-S; mutation in subunit TmoA, 11fold increase in activity with methyl p-tolyl sulfide
-
I100L
-
mutation in subunit TmoA, 0.9fold decrease in activity with styrene; mutation in subunit TmoA, produces 3-methoxycatechol from o-methoxyphenol four times faster than wild-type
-
I100S
-
mutant hydroxylates o-tyrosol, m-tyrosol and p-tyrosol to form hydroxytyrosol; mutation improves both reaction rate and enantioselectivity
-
I100V
-
mutant hydroxylates m-tyrosol to form hydroxytyrosol; mutation improves both reaction rate and enantioselectivity
-
Q141C
-
decrease in regiospecificity for p-cresol formation, mutant functions predominantly as an aromatic ring hydroxylase during the oxidation of p-xylene
-
S395C
-
mutation in subunit TmoA, shows a 15fold increase in 2-phenylethanol hydroxylation rate
-
T201A
-
mutant retains catalytic activity and exhibits 80-90% coupling efficiency compared to 94% for wild-type, with p-cresol representing 90-95% of the total product distribution; mutation has no impact on steady-state catalysis or coupling. Mutant T201A gives stoichometric release of H2O2 during reaction in the absence of substrate and has a faster first-order rate constant for product formation than wild-type; parameters similar to wild-type
-
T201F
-
mutation causes a substantial shift in the product distribution, and gives o- and p-cresol in a 1:1 ratio
-
T201G
-
mutant retains catalytic activity and exhibits 80-90% coupling efficiency compared to 94% for wild-type, with p-cresol representing 90-95% of the total product distribution; mutation has no impact on steady-state catalysis or coupling
-
T201K
-
decrease in activity
-
T201L
-
parameters similar to wild-type
-
T201S
-
mutant retains catalytic activity and exhibits 80-90% coupling efficiency compared to 94% for wild-type, with p-cresol representing 90-95% of the total product distribution; mutation has no impact on steady-state catalysis or coupling; parameters similar to wild-type
-
additional information