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2,4-dichlorophenol + FADH2 + O2
2,4-dichlorocatechol + FAD + H2O
2-aminophenol + O2 + NADPH
?
-
-
-
-
?
2-chlorophenol + FADH2 + O2
2-chlorocatechol + FAD + H2O
2-chlorophenol + O2 + NADPH
?
2-methyl-phenol + O2 + NADPH
?
-
-
-
-
?
3-aminophenol + O2 + NADPH
?
-
-
-
-
?
3-chlorophenol + O2 + NADPH
?
3-methylphenol + O2 + NADPH
?
-
-
-
-
?
3-nitrophenol + NAD(P)H + H+ + O2
? + NAD(P)+ + H2O
4-aminophenol + O2 + NADPH
?
-
-
-
-
?
4-chlorophenol + FADH2 + O2
4-chlorocatechol + FAD + H2O
4-chlorophenol + O2 + NADPH
?
4-methyl-phenol + O2 + NADPH
?
-
-
-
-
?
4-methylphenol + FADH2 + O2
4-methylcatechol + FAD + H2O
-
-
-
?
4-nitrophenol + NAD(P)H + H+ + O2
? + NAD(P)+ + H2O
catechol + FMN + H2O
phenol + FMNH2 + O2
-
-
-
r
catechol + riboflavin + H2O
phenol + reduced riboflavin + O2
-
-
-
r
phenol + FADH2 + O2
catechol + FAD + H2O
phenol + FMNH2 + O2
catechol + FMN + H2O
phenol + NAD(P)H + H+ + O2
catechol + NAD(P)+ + H2O
phenol + NADH + H+ + O2
catechol + NAD+ + H2O
phenol + NADPH + O2
?
-
first step of phenol degradation
-
-
?
phenol + NADPH + O2
catechol + NADP+ + H2O
phenol + reduced riboflavin + O2
catechol + riboflavin + H2O
phloroglucinol + O2 + NADPH
?
-
-
-
-
?
pyrogallol + O2 + NADPH
?
-
-
-
-
?
quinol + O2 + NADPH
1,2,4-trihydroxybenzene + NADP+ + H2O
-
-
-
-
?
resorcinol + NAD(P)H + H+ + O2
? + NAD(P)+ + H2O
resorcinol + NADPH + O2
?
-
-
-
-
?
additional information
?
-
2,4-dichlorophenol + FADH2 + O2
2,4-dichlorocatechol + FAD + H2O
-
-
-
?
2,4-dichlorophenol + FADH2 + O2
2,4-dichlorocatechol + FAD + H2O
-
-
-
?
2-chlorophenol + FADH2 + O2
2-chlorocatechol + FAD + H2O
-
-
-
?
2-chlorophenol + FADH2 + O2
2-chlorocatechol + FAD + H2O
poor substrate for isozyme PheA1(1)
-
-
?
2-chlorophenol + FADH2 + O2
2-chlorocatechol + FAD + H2O
-
-
-
?
2-chlorophenol + FADH2 + O2
2-chlorocatechol + FAD + H2O
poor substrate for isozyme PheA1(1)
-
-
?
2-chlorophenol + O2 + NADPH
?
-
-
-
-
?
2-chlorophenol + O2 + NADPH
?
-
-
-
-
?
3-chlorophenol + O2 + NADPH
?
-
-
-
-
?
3-chlorophenol + O2 + NADPH
?
-
-
-
-
?
3-nitrophenol + NAD(P)H + H+ + O2
? + NAD(P)+ + H2O
-
-
-
?
3-nitrophenol + NAD(P)H + H+ + O2
? + NAD(P)+ + H2O
-
-
-
?
4-chlorophenol + FADH2 + O2
4-chlorocatechol + FAD + H2O
-
-
-
?
4-chlorophenol + FADH2 + O2
4-chlorocatechol + FAD + H2O
-
-
-
?
4-chlorophenol + O2 + NADPH
?
-
-
-
-
?
4-chlorophenol + O2 + NADPH
?
-
-
-
-
?
4-nitrophenol + NAD(P)H + H+ + O2
? + NAD(P)+ + H2O
-
-
-
?
4-nitrophenol + NAD(P)H + H+ + O2
? + NAD(P)+ + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
a two-component enzyme system: the smaller flavin reductase PheA2 component catalyzes the NADH-dependent reduction of free FAD according to a ping pong bisubstrate-biproduct mechanism. The reduced FAD is then used by the larger oxygenase component PheA1 to hydroxylate phenols to the corresponding catechols
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
PheA1 is required for activity, no activity by PheA2 alone. PheA2 acts according to a ping pong bi bi reaction mechanism in which NADH reduces the FAD cofactor, which in turn transfers electrons to the FAD substrate
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
reactive exogenous FAD substrate binds in the NADH cleft after release of NAD product. PheA2 is able to bind one FAD cofactor and one FAD substrate
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
a two-component enzyme system: the smaller flavin reductase PheA2 component catalyzes the NADH-dependent reduction of free FAD according to a ping pong bisubstrate-biproduct mechanism. The reduced FAD is then used by the larger oxygenase component PheA1 to hydroxylate phenols to the corresponding catechols
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
reactive exogenous FAD substrate binds in the NADH cleft after release of NAD product. PheA2 is able to bind one FAD cofactor and one FAD substrate
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
PheA1 is required for activity, no activity by PheA2 alone. PheA2 acts according to a ping pong bi bi reaction mechanism in which NADH reduces the FAD cofactor, which in turn transfers electrons to the FAD substrate
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
hydroxylation of phenol in vitro requires the presence of both PheA1 and PheA2 components, in addition to NADH and FAD, but the physical interaction between the proteins is not necessary for the reaction, Km for FAD is 0.0134 mM, Km for NADH is 0.0533 mM. The hydroxylation of phenol in vitro depends on the molar ratio of His6PheA2 and His6PheA1 present in the reaction mixture, an increase of the amount of His6PheA1 in the assay results in a higher phenol hydroxylase activity. In the assay, a reductase/oxygenase molar ratio of 1:10 is used
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
hydroxylation of phenol in vitro requires the presence of both PheA1 and PheA2 components, in addition to NADH and FAD, but the physical interaction between the proteins is not necessary for the reaction. The hydroxylation of phenol in vitro depends on the molar ratio of His6PheA2 and His6PheA1 present in the reaction mixture, an increase of the amount of His6PheA1 in the assay results in a higher phenol hydroxylase activity. In the assay, a reductase/oxygenase molar ratio of 1:10 is used
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FMNH2 + O2
catechol + FMN + H2O
-
-
-
?
phenol + FMNH2 + O2
catechol + FMN + H2O
-
-
-
?
phenol + NAD(P)H + H+ + O2
catechol + NAD(P)+ + H2O
-
-
-
-
?
phenol + NAD(P)H + H+ + O2
catechol + NAD(P)+ + H2O
hydroxylation of phenol in vitro requires the presence of both His6PheA1 and His6PheA2 components, in addition to NADH and FAD, but the physical interaction between the proteins is not necessary for the reaction
-
-
?
phenol + NAD(P)H + H+ + O2
catechol + NAD(P)+ + H2O
hydroxylation of phenol in vitro requires the presence of both His6PheA1 and His6PheA2 components, in addition to NADH and FAD, but the physical interaction between the proteins is not necessary for the reaction
-
-
?
phenol + NADH + H+ + O2
catechol + NAD+ + H2O
the two-protein system phenol hydroxylase consists of an oxygenase (PheA1) and a flavin reductase (PheA2). PheA1 catalyzes the efficient ortho-hydroxylation of phenol to catechol when supplemented with PheA2 and FAD/NADH. PheA1 catalyzes the NADH-dependent reduction of free flavins according to a ping pong bi bi mechanism
-
-
?
phenol + NADH + H+ + O2
catechol + NAD+ + H2O
the two-protein system phenol hydroxylase consists of an oxygenase (PheA1) and a flavin reductase (PheA2). PheA1 catalyzes the efficient ortho-hydroxylation of phenol to catechol when supplemented with PheA2 and FAD/NADH. PheA1 catalyzes the NADH-dependent reduction of free flavins according to a ping pong bi bi mechanism
-
-
?
phenol + NADH + H+ + O2
catechol + NAD+ + H2O
-
-
-
r
phenol + NADH + H+ + O2
catechol + NAD+ + H2O
-
-
-
r
phenol + NADPH + O2
catechol + NADP+ + H2O
-
-
-
-
?
phenol + NADPH + O2
catechol + NADP+ + H2O
-
-
-
-
?
phenol + reduced riboflavin + O2
catechol + riboflavin + H2O
-
-
-
?
phenol + reduced riboflavin + O2
catechol + riboflavin + H2O
-
-
-
?
resorcinol + NAD(P)H + H+ + O2
? + NAD(P)+ + H2O
-
-
-
?
resorcinol + NAD(P)H + H+ + O2
? + NAD(P)+ + H2O
-
-
-
?
additional information
?
-
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
-
PheA1 is required for activity of flavin reductase PheA2, no activity by PheA1 or PheA2 alone
-
-
?
additional information
?
-
PheA1 is required for activity of flavin reductase PheA2, no activity by PheA1 or PheA2 alone
-
-
?
additional information
?
-
PheA1 is required for activity of flavin reductase PheA2, no activity by PheA1 or PheA2 alone
-
-
?
additional information
?
-
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
-
PheA1 is required for activity of flavin reductase PheA2, no activity by PheA1 or PheA2 alone
-
-
?
additional information
?
-
PheA1 is required for activity of flavin reductase PheA2, no activity by PheA1 or PheA2 alone
-
-
?
additional information
?
-
PheA1 is required for activity of flavin reductase PheA2, no activity by PheA1 or PheA2 alone
-
-
?
additional information
?
-
the two-component phenol hydroxylase is completely unable to hydroxylate benzoate, 4-hydroxybenzoate, and orcinol
-
-
?
additional information
?
-
the two-component phenol hydroxylase is completely unable to hydroxylate benzoate, 4-hydroxybenzoate, and orcinol
-
-
?
additional information
?
-
the two-component phenol hydroxylase is completely unable to hydroxylate benzoate, 4-hydroxybenzoate, and orcinol
-
-
?
additional information
?
-
the two-component phenol hydroxylase is completely unable to hydroxylate benzoate, 4-hydroxybenzoate, and orcinol
-
-
?
additional information
?
-
-
the two-component phenol hydroxylase is completely unable to hydroxylate benzoate, 4-hydroxybenzoate, and orcinol
-
-
?
additional information
?
-
substrate speccificities of the three isozymes, overview
-
-
?
additional information
?
-
substrate speccificities of the three isozymes, overview
-
-
?
additional information
?
-
-
not: 3-nitrophenol, 4-nitrophenol
-
-
?
additional information
?
-
-
not: 3-nitrophenol, 4-nitrophenol
-
-
?
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2,4-dichlorophenol + FADH2 + O2
2,4-dichlorocatechol + FAD + H2O
2-chlorophenol + FADH2 + O2
2-chlorocatechol + FAD + H2O
4-chlorophenol + FADH2 + O2
4-chlorocatechol + FAD + H2O
4-methylphenol + FADH2 + O2
4-methylcatechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
phenol + NADPH + O2
?
-
first step of phenol degradation
-
-
?
additional information
?
-
2,4-dichlorophenol + FADH2 + O2
2,4-dichlorocatechol + FAD + H2O
-
-
-
?
2,4-dichlorophenol + FADH2 + O2
2,4-dichlorocatechol + FAD + H2O
-
-
-
?
2-chlorophenol + FADH2 + O2
2-chlorocatechol + FAD + H2O
poor substrate for isozyme PheA1(1)
-
-
?
2-chlorophenol + FADH2 + O2
2-chlorocatechol + FAD + H2O
poor substrate for isozyme PheA1(1)
-
-
?
4-chlorophenol + FADH2 + O2
4-chlorocatechol + FAD + H2O
-
-
-
?
4-chlorophenol + FADH2 + O2
4-chlorocatechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
a two-component enzyme system: the smaller flavin reductase PheA2 component catalyzes the NADH-dependent reduction of free FAD according to a ping pong bisubstrate-biproduct mechanism. The reduced FAD is then used by the larger oxygenase component PheA1 to hydroxylate phenols to the corresponding catechols
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
a two-component enzyme system: the smaller flavin reductase PheA2 component catalyzes the NADH-dependent reduction of free FAD according to a ping pong bisubstrate-biproduct mechanism. The reduced FAD is then used by the larger oxygenase component PheA1 to hydroxylate phenols to the corresponding catechols
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
phenol + FADH2 + O2
catechol + FAD + H2O
-
-
-
?
additional information
?
-
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
enzyme components PheA2 and PheA1 show no protein-protein interaction
-
-
?
additional information
?
-
substrate speccificities of the three isozymes, overview
-
-
?
additional information
?
-
substrate speccificities of the three isozymes, overview
-
-
?
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FAD
-
FAD
PheA2 uses FAD both as a substrate and as a prosthetic group, strictly dependent on, neither FMN nor riboflavin can replace FAD in this reaction. PheA2 is a a homodimer, with each subunit containing a highly fluorescent FAD prosthetic group
FAD
activity of the oxygenase component His6PheA1 of phenol hydroxylase is strictly dependent on FAD
FADH2
-
FADH2
FAD is again reduced at the expense of NADH and NADPH
FADH2
FAD is bound to PheA2, binding structure analysis, overview
FADH2
-
PheA2 is a single domain homodimeric protein with each FAD-containing subunit being organized around a six-stranded beta-sheet and a capping alpha-helix. The tightly bound FAD prosthetic group binds near the dimer interface, and the re face of the FAD isoalloxazine ring is fully exposed to solvent, binding structure, overview
FMN
-
NADH
-
NADH
PheA2 uses NADH in order to reduce FAD, according to a random sequential kinetic mechanism
NADH
-
addition of NADH to crystalline PheA2 reduces the flavin cofactor, the NAD product is bound in a wide solvent-accessible groove adopting an unusual folded conformation with ring stacking, binding structure, overview
NADH
NADH is a much better coenzyme for PheA2 than NADPH
NADH
preferred compared to NADPH
NADPH
-
-
NADPH
can be used instead of NADH as electron donor, using either FAD or FMN as electron acceptor, but with an affinity 5fold or 10fold lower than NADH, respectively
additional information
-
reactive exogenous FAD substrate binds in the NADH cleft after release of NAD product. PheA2 is able to bind one FAD cofactor and one FAD substrate. PheA2 contains a dual binding cleft for NADH and FAD substrate, which alternate during catalysis. No activity with FMN, riboflavin, and NADPH
-
additional information
the flavoprotein monooxygenase uses electrons of NAD(P)H to activate and cleave a molecule of oxygen through the formation of an intermediate flavin hydroperoxide and enable the incorporation of an oxygen atom into the substrate
-
additional information
the flavoprotein monooxygenase uses electrons of NAD(P)H to activate and cleave a molecule of oxygen through the formation of an intermediate flavin hydroperoxide and enable the incorporation of an oxygen atom into the substrate
-
additional information
-
the free FAD acts as a true substrate. In addition to FAD, PheA2 is also active with FMN and riboflavin. The turnover rate of PheA2 with free flavins is strongly dependent on temperature. At 25°C, the activity with FMN and riboflavin is much higher than with FAD, but when the temperature is raised to 53°C, the turnover rates with the different flavins becomes nearly identical. Dichlorophenolindophenol is a poor cofactor
-
additional information
the free FAD acts as a true substrate. In addition to FAD, PheA2 is also active with FMN and riboflavin. The turnover rate of PheA2 with free flavins is strongly dependent on temperature. At 25°C, the activity with FMN and riboflavin is much higher than with FAD, but when the temperature is raised to 53°C, the turnover rates with the different flavins becomes nearly identical. Dichlorophenolindophenol is a poor cofactor
-
additional information
the free FAD acts as a true substrate. In addition to FAD, PheA2 is also active with FMN and riboflavin. The turnover rate of PheA2 with free flavins is strongly dependent on temperature. At 25°C, the activity with FMN and riboflavin is much higher than with FAD, but when the temperature is raised to 53°C, the turnover rates with the different flavins becomes nearly identical. Dichlorophenolindophenol is a poor cofactor
-
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evolution
phylogenetic analysis shows exceptional high similarities of PheA1(1-3) and PheA2(1-3) to putative phenol hydroxylases in several Rhodococcus strains, overview
evolution
-
phylogenetic analysis shows exceptional high similarities of PheA1(1-3) and PheA2(1-3) to putative phenol hydroxylases in several Rhodococcus strains, overview
-
metabolism
the gene cluster comprising genes catA, catB, catC, catR, pheR, pheA2, pheA1, is involved in the ortho-cleavage pathway of phenol, the key enzyme of the phenol degradation pathway is the two-component phenol hydroxylase. Regulation analysis, the type of carbon catabolite repression and the temporal transcriptional pattern during cultivation are different in each of the analyzed phe clusters from Rhodococcus erythropolis and Rhodococcus jostii
metabolism
the gene cluster comprising genes catA, catB, catC, catR, pheR, pheA2, pheA1, is involved in the ortho-cleavage pathway of phenol, the key enzyme of the phenol degradation pathway is the two-component phenol hydroxylase. Regulation analysis, the type of carbon catabolite repression and the temporal transcriptional pattern during cultivation are different in each of the analyzed phe clusters from Rhodococcus erythropolis and Rhodococcus jostii
metabolism
-
the gene cluster comprising genes catA, catB, catC, catR, pheR, pheA2, pheA1, is involved in the ortho-cleavage pathway of phenol, the key enzyme of the phenol degradation pathway is the two-component phenol hydroxylase. Regulation analysis, the type of carbon catabolite repression and the temporal transcriptional pattern during cultivation are different in each of the analyzed phe clusters from Rhodococcus erythropolis and Rhodococcus jostii
-
physiological function
Geobacillus thermoglucosidasius strain A7 degrades phenol at 65°C via the meta cleavage pathway
physiological function
phenol hydroxylase (PheA) catalyzes the first step in the degradation of phenol in Geobacillus thermoglucosidasius strain A7 is described. The two-protein system, encoded by the pheA1 and pheA2 genes, consists of an oxygenase (PheA1) and a flavin reductase (PheA2)
physiological function
phenol hydroxylase is a two-component enzyme encoded by pheA1 and pheA2 genes and strictly dependent on NADH and FAD. The intracellular enzyme is a clear example of a two-component tetrameric flavoprotein hydroxylase, in which flavin reduction and substrate oxygenation take place in the same protein
physiological function
phenol-degrading aerobic bacteria are able to convert phenol into nontoxic intermediates of the tricarboxylic acid cycle via an ortho or meta pathway. The monooxygenation of the aromatic ring constitutes the first step in the biodegradation of many phenolic compounds. The two-component flavin-dependent monooxygenase phenol hydroxylase catalyzes the conversion of phenol to catechol in Rhodococcus erythropolis UPV-1. Recombinant PheA1 has no phenol hydroxylase activity on its own. Recombinant PheA2 is a flavin reductase that uses NAD(P)H in order to reduce flavin adenine dinucleotide (FAD), according to a random sequential kinetic mechanism. The hydroxylation of phenol in vitro requires the presence of both PheA1 and PheA2 components, in addition to NADH and FAD, but the physical interaction between the proteins is not necessary for the reaction. The enzymic activity catalyzed in vitro by His6PheA2 is essential to carry out the hydroxylation of phenol by His6PheA1
physiological function
-
the catabolism of toxic phenols in the thermophilic organism Bacillus thermoglucosidasius A7 is initiated by a two-component enzyme system. The smaller flavin reductase PheA2 component catalyzes the NADH-dependent reduction of free FAD according to a pingpong bisubstrate-biproduct mechanism. The reduced FAD is then used by the larger oxygenase component PheA1 to hydroxylate phenols to the corresponding catechols
physiological function
-
phenol-degrading aerobic bacteria are able to convert phenol into nontoxic intermediates of the tricarboxylic acid cycle via an ortho or meta pathway. The monooxygenation of the aromatic ring constitutes the first step in the biodegradation of many phenolic compounds. The two-component flavin-dependent monooxygenase phenol hydroxylase catalyzes the conversion of phenol to catechol in Rhodococcus erythropolis UPV-1. Recombinant PheA1 has no phenol hydroxylase activity on its own. Recombinant PheA2 is a flavin reductase that uses NAD(P)H in order to reduce flavin adenine dinucleotide (FAD), according to a random sequential kinetic mechanism. The hydroxylation of phenol in vitro requires the presence of both PheA1 and PheA2 components, in addition to NADH and FAD, but the physical interaction between the proteins is not necessary for the reaction. The enzymic activity catalyzed in vitro by His6PheA2 is essential to carry out the hydroxylation of phenol by His6PheA1
-
physiological function
-
the catabolism of toxic phenols in the thermophilic organism Bacillus thermoglucosidasius A7 is initiated by a two-component enzyme system. The smaller flavin reductase PheA2 component catalyzes the NADH-dependent reduction of free FAD according to a pingpong bisubstrate-biproduct mechanism. The reduced FAD is then used by the larger oxygenase component PheA1 to hydroxylate phenols to the corresponding catechols
-
physiological function
-
phenol hydroxylase is a two-component enzyme encoded by pheA1 and pheA2 genes and strictly dependent on NADH and FAD. The intracellular enzyme is a clear example of a two-component tetrameric flavoprotein hydroxylase, in which flavin reduction and substrate oxygenation take place in the same protein
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physiological function
-
phenol hydroxylase (PheA) catalyzes the first step in the degradation of phenol in Geobacillus thermoglucosidasius strain A7 is described. The two-protein system, encoded by the pheA1 and pheA2 genes, consists of an oxygenase (PheA1) and a flavin reductase (PheA2)
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physiological function
-
Geobacillus thermoglucosidasius strain A7 degrades phenol at 65°C via the meta cleavage pathway
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additional information
-
PheA2 homology modeling using the structure of ferric reductase (FeR) from Archaeoglobus fulgidus with NADP+ bound, PDB ID 1IOS, as template, overview
additional information
PheA2 homology modeling using the structure of ferric reductase (FeR) from Archaeoglobus fulgidus with NADP+ bound, PDB ID 1IOS, as template, overview
additional information
PheA2 homology modeling using the structure of ferric reductase (FeR) from Archaeoglobus fulgidus with NADP+ bound, PDB ID 1IOS, as template, overview
additional information
-
structure analysis and modeling
additional information
-
structure analysis and modeling
-
additional information
-
PheA2 homology modeling using the structure of ferric reductase (FeR) from Archaeoglobus fulgidus with NADP+ bound, PDB ID 1IOS, as template, overview
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16900
-
x * 44200, sequence analysis, monooxygenase subunit of the phenol hydroxylase (PheA1). x * 16900, sequence analysis, flavin reductase subunit of the phenol hydroxylase (PheA2)
17800
2 * 17800, PheA2, about, sequence calculation
238000
non-denaturing-PAGE followed by staining with Coomassie Brilliant Blue
290000 - 310000
isozymes, gel filtration
35000
recombinant PheA2 protein, gel filtration
44200
-
x * 44200, sequence analysis, monooxygenase subunit of the phenol hydroxylase (PheA1). x * 16900, sequence analysis, flavin reductase subunit of the phenol hydroxylase (PheA2)
18000
the oxygenase component PheA1 is a dimer: 2 * 57000, SDS-PAGE. The flavin reductase PheA2 is a dimer: 2 * 18000, SDS-PAGE
18000
2 * 18000, about, recombinant PheA2 protein, SDS-PAGE
18000
x * 18000, about, sequence calculation
20350
2 * 20350, sequence analysis, 2 * 22000, SDS-PAGE, 2 * 22550, mass spectrometry
20350
2 * 22550, recombinant His6-tagged PheA2, mass spectrometry, 2 * 20350, sequence calculation, 2 * 22000, recombinant His6-tagged PheA2, SDS-PAGE
22000
2 * 20350, sequence analysis, 2 * 22000, SDS-PAGE, 2 * 22550, mass spectrometry
22000
2 * 22550, recombinant His6-tagged PheA2, mass spectrometry, 2 * 20350, sequence calculation, 2 * 22000, recombinant His6-tagged PheA2, SDS-PAGE
22550
2 * 20350, sequence analysis, 2 * 22000, SDS-PAGE, 2 * 22550, mass spectrometry
22550
2 * 22550, recombinant His6-tagged PheA2, mass spectrometry, 2 * 20350, sequence calculation, 2 * 22000, recombinant His6-tagged PheA2, SDS-PAGE
236000
gel filtration
236000
recombinant PheA1, gel filtration
45000
gel filtration
45000
recombinant PheA2, gel filtration
57000
2 * 57000, SDS-PAGE
57000
the oxygenase component PheA1 is a dimer: 2 * 57000, SDS-PAGE. The flavin reductase PheA2 is a dimer: 2 * 18000, SDS-PAGE
57000
x * 57000, about, sequence calculation
60720
4 * 60720, sequence analysis, 4 * 62000, SDS-PAGE, 4 * 62078, mass spectrometry
60720
4 * 62078, recombinant His6-tagged PheA1, mass spectrometry, 4 * 60720, sequence calculation, 4 * 62000, recombinant His6-tagged PheA1, SDS-PAGE
62000
4 * 60720, sequence analysis, 4 * 62000, SDS-PAGE, 4 * 62078, mass spectrometry
62000
4 * 62078, recombinant His6-tagged PheA1, mass spectrometry, 4 * 60720, sequence calculation, 4 * 62000, recombinant His6-tagged PheA1, SDS-PAGE
62078
4 * 60720, sequence analysis, 4 * 62000, SDS-PAGE, 4 * 62078, mass spectrometry
62078
4 * 62078, recombinant His6-tagged PheA1, mass spectrometry, 4 * 60720, sequence calculation, 4 * 62000, recombinant His6-tagged PheA1, SDS-PAGE
additional information
-
PheA1 has a MW of 120000 Da as determined by gel filtration. PheA2 has a MW of 35000 Da as determined by gel filtration
additional information
PheA1 has a MW of 120000 Da as determined by gel filtration. PheA2 has a MW of 35000 Da as determined by gel filtration
additional information
PheA1 has a MW of 120000 Da as determined by gel filtration. PheA2 has a MW of 35000 Da as determined by gel filtration
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?
-
x * 44200, sequence analysis, monooxygenase subunit of the phenol hydroxylase (PheA1). x * 16900, sequence analysis, flavin reductase subunit of the phenol hydroxylase (PheA2)
?
x * 18000, about, sequence calculation
?
x * 57000, about, sequence calculation
?
-
x * 18000, about, sequence calculation
-
?
-
x * 57000, about, sequence calculation
-
homodimer
2 * 57000, SDS-PAGE
homodimer
2 * 17800, PheA2, about, sequence calculation
homodimer
2 * 18000, about, recombinant PheA2 protein, SDS-PAGE
homodimer
-
PheA2 is a single domain homodimeric protein with each FAD-containing subunit being organized around a six-stranded beta-sheet and a capping alpha-helix. The tightly bound FAD prosthetic group binds near the dimer interface, and the re face of the FAD isoalloxazine ring is fully exposed to solvent. PheA2 contains a dual binding cleft for NADH and FAD substrate, which alternate during catalysis
homodimer
-
PheA2 is a single domain homodimeric protein with each FAD-containing subunit being organized around a six-stranded beta-sheet and a capping alpha-helix. The tightly bound FAD prosthetic group binds near the dimer interface, and the re face of the FAD isoalloxazine ring is fully exposed to solvent. PheA2 contains a dual binding cleft for NADH and FAD substrate, which alternate during catalysis
-
homodimer
-
2 * 17800, PheA2, about, sequence calculation
-
homodimer
-
2 * 18000, about, recombinant PheA2 protein, SDS-PAGE
-
homodimer
-
2 * 57000, SDS-PAGE
-
homodimer
2 * 20350, sequence analysis, 2 * 22000, SDS-PAGE, 2 * 22550, mass spectrometry
homodimer
2 * 22550, recombinant His6-tagged PheA2, mass spectrometry, 2 * 20350, sequence calculation, 2 * 22000, recombinant His6-tagged PheA2, SDS-PAGE
homodimer
-
2 * 20350, sequence analysis, 2 * 22000, SDS-PAGE, 2 * 22550, mass spectrometry
-
homodimer
-
2 * 22550, recombinant His6-tagged PheA2, mass spectrometry, 2 * 20350, sequence calculation, 2 * 22000, recombinant His6-tagged PheA2, SDS-PAGE
-
homotetramer
4 * 60720, sequence analysis, 4 * 62000, SDS-PAGE, 4 * 62078, mass spectrometry
homotetramer
4 * 62078, recombinant His6-tagged PheA1, mass spectrometry, 4 * 60720, sequence calculation, 4 * 62000, recombinant His6-tagged PheA1, SDS-PAGE
homotetramer
-
4 * 60720, sequence analysis, 4 * 62000, SDS-PAGE, 4 * 62078, mass spectrometry
-
homotetramer
-
4 * 62078, recombinant His6-tagged PheA1, mass spectrometry, 4 * 60720, sequence calculation, 4 * 62000, recombinant His6-tagged PheA1, SDS-PAGE
-
oligomer
homotetrameric or homohexameric structure, all phenol hydroxylase isoenzymes, x * 59000-63000, SDS-PAGE
oligomer
-
homotetrameric or homohexameric structure, all phenol hydroxylase isoenzymes, x * 59000-63000, SDS-PAGE
-
additional information
-
the oxygenase component PheA1 is a dimer: 2 * 57000, SDS-PAGE. The flavin reductase PheA2 is a dimer: 2 * 18000, SDS-PAGE
additional information
the oxygenase component PheA1 is a dimer: 2 * 57000, SDS-PAGE. The flavin reductase PheA2 is a dimer: 2 * 18000, SDS-PAGE
additional information
the oxygenase component PheA1 is a dimer: 2 * 57000, SDS-PAGE. The flavin reductase PheA2 is a dimer: 2 * 18000, SDS-PAGE
additional information
the large subunit of this enzyme is encoded by pheA1 gene and gives rise to a 57 kDa protein. The protein has hydroxylase activity, is a homodimer, but has no activity by its own
additional information
the large subunit of this enzyme is encoded by pheA1 gene and gives rise to a 57 kDa protein. The protein has hydroxylase activity, is a homodimer, but has no activity by its own
additional information
-
the large subunit of this enzyme is encoded by pheA1 gene and gives rise to a 57 kDa protein. The protein has hydroxylase activity, is a homodimer, but has no activity by its own
additional information
the minor component encoded by pheA2 gene has a predicted molecular mass of 17.8 kDa, it is a homodimer and shows a specific NADH:FAD reductase activity
additional information
the minor component encoded by pheA2 gene has a predicted molecular mass of 17.8 kDa, it is a homodimer and shows a specific NADH:FAD reductase activity
additional information
-
the minor component encoded by pheA2 gene has a predicted molecular mass of 17.8 kDa, it is a homodimer and shows a specific NADH:FAD reductase activity
additional information
-
the oxygenase component PheA1 is a dimer: 2 * 57000, SDS-PAGE. The flavin reductase PheA2 is a dimer: 2 * 18000, SDS-PAGE
-
additional information
-
the minor component encoded by pheA2 gene has a predicted molecular mass of 17.8 kDa, it is a homodimer and shows a specific NADH:FAD reductase activity
-
additional information
-
the large subunit of this enzyme is encoded by pheA1 gene and gives rise to a 57 kDa protein. The protein has hydroxylase activity, is a homodimer, but has no activity by its own
-
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gene pheA1, DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli, cloning of the pheA1 promoter from Geobacillus thermoglucosidasius and use for enzyme expression in Escherichia coli strain Rosetta, tandem expression of genes pheA1 and pheA2
gene pheA1, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression in Escherichia coli, coexpression with pheA2 is required for catalytic activity
gene pheA1, quantitative real-time PCR-based expression and promoter activity analysis, recombinant expression in Corynebacterium glutamicum strain RES167, a heterologous host lacking the PheR regulator, coexpression of gene pheA1 with gene pheA2 and regulator gene pheR, subcloning in Escherichia coli strain DH5alpha
gene pheA1, recombinant expression in Escherichia coli, tandem expression with pheA2 does not result in PheA2 protein
gene pheA2, DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli strain Rosetta tandem expression of genes pheA1 and pheA2
gene pheA2, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression in Escherichia coli, coexpression with pheA1 is required for catalytic activity
gene pheA2, recombinant expression in Escherichia coli, tandem expression with pheA1 does not result in PheA2 protein
gene pheA2, sequence comparisons, recombinant overexpression of wild-type and selenomethionine-substituted PheA2 in Escherichia coli strain BL21(DE3)pLysS
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genes pheA1(1-3), phylogenetic analysis, typical for genes of the peripheral degradation of aromatic compounds, pheA1(13) and pheA2(1-3) are not located within gene clusters for central ortho- and meta-cleavage pathway. All three gene sets are nearby to genes with function in (chloro)aromatic degradation
PheA1 and PheA2 are separately expressed in recombinant Escherichia coli BL21 pLysS cells
phenol hydroxylase is a two-component flavin-dependent monooxygenase, the two proteins are encoded by the genes pheA1 and pheA2, located very closely in the genome, DNA and amino acid sequence determination and analysis, recombinant expression of His6-tagged PheA1 in Escherichia coli strain M15 by nickel affinity chromatography
phenol hydroxylase is a two-component flavin-dependent monooxygenase, the two proteins are encoded by the genes pheA1 and pheA2, located very closely in the genome, DNA and amino acid sequence determination and analysis, recombinant expression of His6-tagged PheA2 in Escherichia coli strain M15
plasmid pQE30A2 expressing His6PheA2 protein transformed into Escherichia coli M15
plasmid pQE9A1 expressing His6PheA1 protein transformed into Escherichia coli M15
gene pheA1, quantitative real-time PCR-based expression and promoter activity analysis, recombinant expression in Corynebacterium glutamicum strain RES167, a heterologous host lacking the PheR regulator, coexpression of gene pheA1 with gene pheA2 and regulator gene pheR, subcloning in Escherichia coli strain DH5alpha
gene pheA1, quantitative real-time PCR-based expression and promoter activity analysis, recombinant expression in Corynebacterium glutamicum strain RES167, a heterologous host lacking the PheR regulator, coexpression of gene pheA1 with gene pheA2 and regulator gene pheR, subcloning in Escherichia coli strain DH5alpha
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Straube, G.
Phenol hydroxylase from Rhodococcus sp. P 1
J. Basic Microbiol.
27
229-232
1987
Rhodococcus sp., Rhodococcus sp. P1
brenda
Kirchner, U.; Westphal, A.H.; Muller, R.; van Berkel, W.J.
Phenol hydroxylase from Bacillus thermoglucosidasius A7, a two-protein component monooxygenase with a dual role for FAD
J. Biol. Chem.
278
47545-47553
2003
Parageobacillus thermoglucosidasius, Parageobacillus thermoglucosidasius (Q9LAG2), Parageobacillus thermoglucosidasius (Q9LAG3), Parageobacillus thermoglucosidasius A7, Parageobacillus thermoglucosidasius A7 (Q9LAG2), Parageobacillus thermoglucosidasius A7 (Q9LAG3)
brenda
Omokoko, B.; Jaentges, U.K.; Zimmermann, M.; Reiss, M.; Hartmeier, W.
Isolation of the phe-operon from G. stearothermophilus comprising the phenol degradative meta-pathway genes and a novel transcriptional regulator
BMC Microbiol.
8
197
2008
Geobacillus stearothermophilus
brenda
Saa, L.; Jaureguibeitia, A.; Largo, E.; Llama, M.J.; Serra, J.L.
Cloning, purification and characterization of two components of phenol hydroxylase from Rhodococcus erythropolis UPV-1
Appl. Microbiol. Biotechnol.
86
201-211
2009
Rhodococcus erythropolis (A7LCL0), Rhodococcus erythropolis (A7LCL1), Rhodococcus erythropolis UPV-1 (A7LCL0), Rhodococcus erythropolis UPV-1 (A7LCL1), Rhodococcus erythropolis UPV-1
brenda
Szoekoel, J.; Rucka, L.; Simcikova, M.; Halada, P.; Nesvera, J.; Patek, M.
Induction and carbon catabolite repression of phenol degradation genes in Rhodococcus erythropolis and Rhodococcus jostii
Appl. Microbiol. Biotechnol.
98
8267-8279
2014
Rhodococcus erythropolis (A7LCL0), Rhodococcus erythropolis, Rhodococcus jostii (Q0SDR7), Rhodococcus jostii, Rhodococcus erythropolis CCM2595 (A7LCL0), Rhodococcus erythropolis CCM2595
brenda
Groening, J.; Eulberg, D.; Tischler, D.; Kaschabek, S.; Schloemann, M.
Gene redundancy of two-component (chloro)phenol hydroxylases in Rhodococcus opacus 1CP
FEMS Microbiol. Lett.
361
68-75
2014
Rhodococcus opacus (A0A069AW73), Rhodococcus opacus 1CP (A0A069AW73)
brenda
Orenes-Pinero, E.; Garcia-Carmona, F.; Sanchez-Ferrer, A.
A new process for obtaining hydroxytyrosol using transformed Escherichia coli whole cells with phenol hydroxylase gene from Geobacillus thermoglucosidasius
Food Chem.
139
377-383
2013
Parageobacillus thermoglucosidasius (Q9LAG2), Parageobacillus thermoglucosidasius (Q9LAG3), Parageobacillus thermoglucosidasius, Parageobacillus thermoglucosidasius A7 (Q9LAG2), Parageobacillus thermoglucosidasius A7 (Q9LAG3)
brenda
Duffner, F.M.; Kirchner, U.; Bauer, M.P.; Mueller, R.
Phenol/cresol degradation by the thermophilic Bacillus thermoglucosidasius A7: cloning and sequence analysis of five genes involved in the pathway
Gene
256
215-221
2000
Parageobacillus thermoglucosidasius (Q9LAG2), Parageobacillus thermoglucosidasius (Q9LAG3), Parageobacillus thermoglucosidasius, Parageobacillus thermoglucosidasius A7 (Q9LAG2), Parageobacillus thermoglucosidasius A7 (Q9LAG3)
brenda
van den Heuvel, R.H.; Westphal, A.H.; Heck, A.J.; Walsh, M.A.; Rovida, S.; van Berkel, W.J.; Mattevi, A.
Structural studies on flavin reductase PheA2 reveal binding of NAD in an unusual folded conformation and support novel mechanism of action
J. Biol. Chem.
279
12860-12867
2004
Parageobacillus thermoglucosidasius, Parageobacillus thermoglucosidasius A7
brenda