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(2-methylphenyl)acetone + NADPH + O2
? + NADP+ + O2
-
-
-
-
?
(2R,3S)-3-methyl-2-pentylcyclopentanone + NADPH + H+ + O2
?
-
less than 5% conversion
-
-
?
(R)-1-acetoxy-phenylacetone + NADPH + O2
(R)-1-hydroxy-1-phenylacetone + NADP+ + H2O
-
-
-
-
?
(R)-2-acetoxyphenylacetonitrile + NADPH + H+ + O2
?
-
enantioselective reaction
-
-
?
(R)-3-(4-bromophenyl)butan-2-one + NADPH + H+ + O2
?
-
enantioselective reaction
-
-
?
(S)-1-(3-trifluoromethylphenyl)ethyl acetate + NADPH + H+ + O2
?
-
enantioselective reaction
-
-
?
(S)-nicotine + NADPH + O2
?
-
-
-
-
?
1-bromo-indanone + NADPH + O2
6-bromoisochroman-1-one + NADP+ + H2O
-
substrate was only accepted by phenylacetone monooxygenase of Pseudomonas fluorescens but not of Thermobifida fusca, reaction was performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
1-indanone + NADPH + H+ + O2
1-isochromanone + NADP+ + H2O
-
reaction product is only synthesized by the mutant M446G of phenylacetone monooxygenase of Thermobifida fusca, reaction is performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
1-indanone + NADPH + H+ + O2
3,4-dihydrocoumarin + NADP+ + H2O
-
substrate was only accepted by phenylacetone monooxygenase of Pseudomonas fluorescens but not of Thermobifida fusca, reaction was performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
1-tetralone + NADPH + O2
4,5-dihydro-1-benzoxepin-2(3H)-one + NADP+ + H2O
-
substrate was accepted by phenylacetone monooxygenase of Pseudomonas fluorescens but not of Thermobifida fusca, reaction was performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
1-[3-(benzylselanyl)phenyl]ethanone + NADPH + H+ + O2
1-(3-(benzylseleninyl)phenyl)ethanone + NADP+ + H2O
-
more than 99% conversion
-
-
?
1-[3-(methylselanyl)phenyl]ethanone + NADPH + H+ + O2
1-(3-(methylseleninyl)phenyl)ethanone + NADP+ + H2O
-
76% conversion
-
-
?
1-[4-(benzylselanyl)phenyl]ethanone + NADPH + H+ + O2
1-(4-(benzylseleninyl)phenyl)ethanone + NADP+ + H2O
-
more than 99% conversion
-
-
?
1-[4-(methylselanyl)phenyl]ethanone + NADPH + H+ + O2
1-(4-(methylseleninyl)phenyl)ethanone + NADP+ + H2O
-
more than 99% conversion
-
-
?
2-benzylcyclopentanone + NADPH + H+ + O2
?
-
about 10% conversion
-
-
?
2-decanone + NADPH + O2
? + NADP+ + H2O
2-decanone + NADPH + O2
methyl nonanoate + octyl acetate + NADP+ + H2O
-
-
-
-
?
2-dodecanone + NADPH + O2
? + NADP+
-
-
-
-
?
2-dodecanone + NADPH + O2
? + NADP+ + H2O
2-dodecanone + NADPH + O2
nonyl acetate + methyl decanoate + NADP+ + H2O
-
-
-
-
?
2-heptanone + NADPH + O2
pentyl acetate + NADP+ + H2O
-
-
-
-
?
2-hexanone + NADPH + O2
butyl acetate + NADP+ + H2O
-
-
-
-
?
2-indanone + NADPH + H+ + O2
3-isochromanone + NADP+ + H2O
-
substrate is only accepted by the mutant M446G of phenylacetone monooxygenase of Thermobifida fusca, reaction is performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
2-methylcyclohexanone + NADPH + H+ + O2
? + NADP+ + H2O
no activity with wild-type PAMO, but low to increased activity with enzyme mutants
-
-
?
2-methylphenylcyclohexanone + NADPH + O2
7-benzyloxepan-2-one + NADP+ + H2O
-
mutant P3 prefers the R-isomer
-
-
?
2-nonanone + NADPH + H+ + O2
?
-
less than 40% conversion
-
-
?
2-nonanone + NADPH + O2
? + NADP+ + H2O
2-octanone + NADPH + H+ + O2
?
-
-
-
-
?
2-octanone + NADPH + H+ + O2
? + NADP+ + H2O
-
-
-
?
2-octanone + NADPH + H+ + O2
heptylacetate + NADP+ + H2O
-
-
-
?
2-octanone + NADPH + O2
heptyl acetate + NADP+ + H2O
-
-
-
-
?
2-phenylcyclohexanone + NADPH + H+ + O2
?
-
-
-
-
?
2-phenylcyclohexanone + NADPH + H+ + O2
? + NADP+ + H2O
very low activity with wild-type PAMO, significantly increased activity with enzyme mutant P253F/G254A/R258M/L443F
-
-
?
2-phenylcyclohexanone + NADPH + O2
7-phenyloxepan-2-one + NADP+ + H2O
-
molecular modeling of the Criegee intermediate, the wild-type enzyme prefers the S-isomer, while mutants P1-P3 all prefer the R-isomer
-
-
?
2-phenylpropionaldehyde + NADPH + H+ + O2
?
-
-
-
?
2-undecanone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
3-(3-trifluoromethylphenyl)butan-2-one + NADPH + H+ + O2
?
-
enantioselective reaction
-
-
?
3-(4-chlorophenyl)cyclobutanone + NADPH + H+ + O2
?
-
about 40% conversion
-
-
?
3-benzylcyclobutanone + NADPH + H+ + O2
?
-
about 45% conversion
-
-
?
3-decanone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
3-octanone + NADPH + H+ + O2
?
-
-
-
-
?
3-octanone + NADPH + O2
ethyl hexanoate + pentyl propanoate + NADP+ + H2O
-
-
-
-
?
3-phenyl-2-butanone + NADPH + H+ + O2
(R)-3-phenylbutan-2-one + (S)-1-phenyethyl acetate
-
enantioselective reaction
-
-
?
3-phenylcyclobutanone + NADPH + H+ + O2
?
-
about 70% conversion
-
-
?
3-phenylpenta-2,4-dione + NADPH + O2
(R)-phenylacetylcarbinol + NADP+ + H2O
-
-
the product is a well-known precursor in the synthesis of ephedrine and pseudoephedrine
-
?
4-decanone + NADPH + O2
? + NADP+ + H2O
4-heptanone + NADPH + O2
propanyl butanoate + NADP+ + H2O
-
-
-
-
?
4-hydroxyacetophenone + NADPH + O2
acetic acid 4-hydroxyphenyl ester + NADP+ + O2
-
-
-
-
?
4-methylcyclohexanone + NADPH + H+ + O2
? + NADP+ + H2O
no activity with wild-type PAMO, but low to increased activity with enzyme mutants
-
-
?
4-phenylcyclohexanone + NADPH + O2
4-phenyl-hexano-6-lactone + NADP+ + H2O
-
-
-
-
?
6-methoxy-1-indanone + NADPH + O2
? + NADP+ + H2O
-
substrate was only accepted by phenylacetone monooxygenase of Pseudomonas fluorescens but not of Thermobifida fusca, reaction was performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
alpha-acetylphenylacetonitrile + NADPH + H+ + O2
(R)-2-acetoxyphenylacetonitrile + NADP+ + H2O
-
enantioselective reaction
enantiopure product formation
-
?
benzocyclobutanone + NADPH + O2
2-coumaranone + NADP+ + H2O
benzylacetone + NADPH + O2
?
-
low activity
-
-
?
benzylacetone + NADPH + O2
? + NADP+ + H2O
bicyclohept-2-en-6-one + NADPH + O2
3-oxabicyclo[3.3.0]oct-6-en-2-one + NADP+ + H2O
-
-
-
-
?
bicyclohept-2-en-6-one + NADPH + O2
? + NADP+ + O2
-
-
-
-
?
bicyclo[2.2.1]heptan-2-one + NADPH + H+ + O2
?
-
about 5% conversion
-
-
?
bicyclo[3.2.0]hept-2-en-6-one + NADPH + H+ + O2
?
-
-
-
-
?
cycloheptanone + NADPH + H+ + O2
? + NADP+ + H2O
no activity with wild-type PAMO, but low to increased activity with enzyme mutants
-
-
?
cyclohexanone + NADPH + H+ + O2
?
-
-
-
-
?
cyclohexanone + NADPH + H+ + O2
epsilon-caprolactone + NADP+ + H2O
cyclopentanone + NADPH + H+ + O2
5-valerolactone + NADP+ + H2O
cyclopentanone + NADPH + H+ + O2
?
-
-
-
-
?
diketone + NADPH + O2
(R)-1-acetoxy-phenylacetone + NADP+ + H2O
-
-
-
-
?
ethionamide + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
ethionamide + NADPH + O2
? + NADP+ + O2
-
-
-
-
?
methyl 4-tolylsulfide + NADPH + O2
? + NADP+ + O2
-
-
-
-
?
methyl-p-tolylsulfide + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
N,N-dimethylbenzylamine + NADPH + O2
N,N-dimethylbenzylamine N-oxide + NADP+ + H2O
-
-
-
-
?
phenylacetone + NADPH + H+ + O2
benzyl acetate + NADP+ + H2O
phenylacetone + NADPH + O2
benzyl acetate + NADP+ + H2O
phenylboronic acid + NADPH + O2
?
-
formation of phenol
-
-
?
rac-2-ethylcyclohexanone + NADPH + O2
benzyl acetate + NADP+ + H2O
-
substrate is only accepted by mutants of phenylacetone monooxygenase, reaction is performed in presence of 2 U secondary alcohol dehydrogenase from Thermoanaerobacter ethanolicus and isopropanol to recover NADPH
-
-
?
rac-3-methyl-4-phenylbutan-2-one + NADH + H+ + O2
(2R)-1-phenylpropan-2-yl acetate + NAD+ + H2O
-
enantioselective reaction by PAMO
-
-
?
rac-3-methyl-4-phenylbutan-2-one + NADPH + H+ + O2
(2R)-1-phenylpropan-2-yl acetate + NADP+ + H2O
-
enantioselective reaction by PAMO
-
-
?
rac-bicyclo [3.2.0]hept-2-en-6-one + NADPH + O2
?
-
activity and stereoselectivity of wild-type and mutant enzymes, overview
-
-
?
thioanisole + NADH + H+ + O2
thioanisole sulfoxide + NAD+ + H2O
-
low activity, less enantioselective reaction
-
-
?
thioanisole + NADPH + H+ + O2
?
thioanisole + NADPH + H+ + O2
methyl phenyl sulfoxide + NADP+ + H2O
-
-
-
-
?
thioanisole + NADPH + H+ + O2
thioanisole sulfoxide + NADP+ + H2O
-
enantioselective reaction
mainly (R)-sulfoxide
-
?
additional information
?
-
2-decanone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
2-decanone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
2-dodecanone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
2-dodecanone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
2-nonanone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
2-nonanone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
4-decanone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
4-decanone + NADPH + O2
? + NADP+ + H2O
-
best substrate
-
-
?
benzocyclobutanone + NADPH + O2
2-coumaranone + NADP+ + H2O
-
reaction was performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
benzocyclobutanone + NADPH + O2
2-coumaranone + NADP+ + H2O
-
reaction is performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
benzylacetone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
benzylacetone + NADPH + O2
? + NADP+ + H2O
-
-
-
-
?
cyclohexanone + NADPH + H+ + O2
epsilon-caprolactone + NADP+ + H2O
substrate of enzyme mutants, not of wild-type, overview
-
-
?
cyclohexanone + NADPH + H+ + O2
epsilon-caprolactone + NADP+ + H2O
no activity with wild-type PAMO, but low to increased activity with enzyme mutants
-
-
?
cyclopentanone + NADPH + H+ + O2
5-valerolactone + NADP+ + H2O
-
-
-
?
cyclopentanone + NADPH + H+ + O2
5-valerolactone + NADP+ + H2O
very low activity with wild-type PAMO, significantly increased activity with enzyme mutant P253F/G254A/R258M/L443F
-
-
?
phenylacetone + NADPH + H+ + O2
benzyl acetate + NADP+ + H2O
-
-
-
-
?
phenylacetone + NADPH + H+ + O2
benzyl acetate + NADP+ + H2O
-
-
-
-
?
phenylacetone + NADPH + H+ + O2
benzyl acetate + NADP+ + H2O
-
-
-
?
phenylacetone + NADPH + H+ + O2
benzyl acetate + NADP+ + H2O
-
-
-
-
?
phenylacetone + NADPH + H+ + O2
benzyl acetate + NADP+ + H2O
-
-
-
?
phenylacetone + NADPH + H+ + O2
benzyl acetate + NADP+ + H2O
-
-
-
-
?
phenylacetone + NADPH + H+ + O2
benzyl acetate + NADP+ + H2O
-
-
-
?
phenylacetone + NADPH + H+ + O2
benzyl acetate + NADP+ + H2O
enantioselective reaction, regulation mechanism, overview
-
-
?
phenylacetone + NADPH + O2
benzyl acetate + NADP+ + H2O
-
reaction was performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
phenylacetone + NADPH + O2
benzyl acetate + NADP+ + H2O
-
reaction is performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH
-
-
?
phenylacetone + NADPH + O2
benzyl acetate + NADP+ + H2O
-
reaction is performed in presence of 2 U secondary alcohol dehydrogenase from Thermoanaerobacter ethanolicus and isopropanol to recover NADPH
-
-
?
thioanisole + NADPH + H+ + O2
?
-
-
-
-
?
thioanisole + NADPH + H+ + O2
?
the asymmetric oxidation of thioanisole to sulfoxide is accompanied by the overoxidation to achiral sulfone
-
-
?
additional information
?
-
-
substrate specificity and reaction mechanism, the enzyme shows high specificity towards short-chain aliphatic ketones, some open-chain ketones are converted to the alkylacetates, while for others formation of the ester products with oxygen on the other side of the keto group can also be detected yielding the corresponding methyl or ethyl esters, overview
-
-
?
additional information
?
-
-
the enzyme shows high specificity towards short-chain aliphatic ketones, some open-chain ketones are converted to the alkylacetates, while for others formation of the ester products with oxygen on the other side of the keto group can also be detected yielding the corresponding methyl or ethyl esters, overview, no activity with 2-octanone and 2-tridecanone
-
-
?
additional information
?
-
-
substrate specificity and reaction mechanism, the enzyme shows high specificity towards short-chain aliphatic ketones, some open-chain ketones are converted to the alkylacetates, while for others formation of the ester products with oxygen on the other side of the keto group can also be detected yielding the corresponding methyl or ethyl esters, overview
-
-
?
additional information
?
-
-
the enzyme shows high specificity towards short-chain aliphatic ketones, some open-chain ketones are converted to the alkylacetates, while for others formation of the ester products with oxygen on the other side of the keto group can also be detected yielding the corresponding methyl or ethyl esters, overview, no activity with 2-octanone and 2-tridecanone
-
-
?
additional information
?
-
substrate specificity of wild-type and mutant enzymes, overview
-
-
-
additional information
?
-
-
enzyme activity in a variety of different aqueousorganic media using organic sulfides as substrates, enantioselectivity, overview
-
-
?
additional information
?
-
-
substrate selectivity and stereospecificity of wild-type and mutant enzymes, overview
-
-
?
additional information
?
-
-
the recombinant His-tagged enzyme is active with a large range of sulfides and ketones, as well as with several sulfoxides, an amine and an organoboron compound, high enantioselectivity dependeing on the substrate, substrate specificity, overview
-
-
?
additional information
?
-
PAMO is an FAD-containing Baeyer-Villiger monooxygenase
-
-
?
additional information
?
-
-
PAMO is an FAD-containing Baeyer-Villiger monooxygenase
-
-
?
additional information
?
-
PAMO is an FAD-containing Baeyer-Villiger monooxygenase, two different residues are responsible for the pH effects on PAMO enantioselectivity, protonation of Arg337 and the FAD:C4a-hydroperoxide/FAD:C4a-peroxide equilibrium are the major factors responsible for the fine-tuning of PAMO enantioselectivity in Baeyer-Villiger oxidation and sulfooxidation, respectively
-
-
?
additional information
?
-
-
determination of enantioselectivity of wild-type and mutant enzymes with different substrates and cofactors, overview. Residue K336 has a significant and beneficial effect on the enantioselectivity of Baeyer-Villiger oxidations and sulfoxidations
-
-
?
additional information
?
-
binding of cyclopentanone and 2-phenylcyclohexanone, which are the typical substrates of CPMO in group I and CHMO in group III, respectively, is analyzed with wild-type and mutant PAMO enzymes. Substrate binding analysis, overview. Residue R337 establishes a cationn-i interaction with the substrate
-
-
-
additional information
?
-
for wild-type TfPAMO, the uncoupling side activity at different pHs ranges from 5.0 to 7.8%. Formation of hydrogen peroxide is rather constant (around 4.0 microM), while formation of superoxide is almost absent at low pH of 6.0, reaching 3.0 microM at high pH of 9.0-10.0
-
-
-
additional information
?
-
the enzyme produces H2O2 in a side uncoupling reaction. The enzyme access tunnels, which may serve as exit paths for H2O2 from the active site to the bulk, are predicted by using the CAVER and/or protein energy landscape exploration (PELE) software for the phenylacetone monooxygenase variant (PAMO_C65D) from Thermobifida fusca. The simplified mechanism of flavin-dependent monooxygenases (e.g. BVMOs) consists of NAD(P)H-dependent reduction of the flavin prosthetic group, followed by activation of molecular oxygen as a (hydro)peroxyflavin, and substrate oxygenation. The catalytic cycle is closed after elimination of water and reformation of the oxidized flavin. Alternatively, the (hydro)peroxyflavin can eliminate H2O2 spontaneously (uncoupling reaction) into the oxidized flavin
-
-
-
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0.83
(2-methylphenyl)acetone
-
30°C, pH 7.5, 0.1 mM NADPH
0.07 - 44
2-phenylcyclohexanone
2.2
4-hydroxyacetophenone
-
30°C, pH 7.5, 0.1 mM NADPH
0.36 - 0.52
benzylacetone
15
bicyclohept-2-en-6-one
-
30°C, pH 7.5, 0.1 mM NADPH
0.1037 - 0.698
cyclohexanone
1000 - 1200
Cyclopentanone
0.34
Ethionamide
-
25°C, pH 8.5
0.86
methyl 4-tolylsulfide
-
30°C, pH 7.5, 0.1 mM NADPH
additional information
additional information
-
0.2
2-dodecanone
-
25°C, pH 8.5
0.26
2-dodecanone
-
30°C, pH 7.5, 0.1 mM NADPH
0.25
2-Octanone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
0.25
2-Octanone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
0.3
2-Octanone
recombinant mutant R258M, pH 7.4, 25°C
2
2-Octanone
-
wild type enzyme, at pH 7.5 and 37°C
2.1
2-Octanone
recombinant mutant R258A, pH 7.4, 25°C
3.2
2-Octanone
recombinant wild-type enzyme, pH 7.4, 25°C
0.07
2-phenylcyclohexanone
-
mutant P3
0.5
2-phenylcyclohexanone
-
mutant P2
0.8
2-phenylcyclohexanone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
0.8
2-phenylcyclohexanone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
2.3
2-phenylcyclohexanone
-
mutant P1
4
2-phenylcyclohexanone
-
wild type enzyme, at pH 7.5 and 37°C
44
2-phenylcyclohexanone
recombinant wild-type enzyme, pH 7.4, 25°C
0.36
benzylacetone
-
30°C, pH 7.5, 0.1 mM NADPH
0.52
benzylacetone
-
25°C, pH 8.5
0.1037
cyclohexanone
mutant P440F, pH 8.0, 45°C
0.2107
cyclohexanone
mutant I67Y/P440F, pH 8.0, 45°C
0.2595
cyclohexanone
mutant I67G/P440F, pH 8.0, 45°C
0.266
cyclohexanone
pH 8.0, 25°C, recombinant mutant S441G/A442P/L443T/S444Q
0.3472
cyclohexanone
mutant I67A/P440F, pH 8.0, 45°C
0.4412
cyclohexanone
mutant I67C/P440F, pH 8.0, 45°C
0.698
cyclohexanone
pH 8.0, 25°C, recombinant mutant A442P/L443V
1000
Cyclopentanone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
1000
Cyclopentanone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
1200
Cyclopentanone
recombinant wild-type enzyme, pH 7.4, 25°C
1200
Cyclopentanone
-
wild type enzyme, at pH 7.5 and 37°C
0.0006
NADPH
-
pH 8.0, 30°C, recombinant mutant H220N
0.0007
NADPH
-
pH 8.0, 30°C, recombinant mutant H220A
0.0007
NADPH
-
pH 8.0, 30°C, recombinant wild-type PAMO
0.0007
NADPH
-
at pH 9.0 and 20°C
0.0011
NADPH
-
pH 8.0, 30°C, recombinant mutant K336N
0.0014
NADPH
-
pH 8.0, 30°C, recombinant mutant T218A
0.0017
NADPH
-
pH 8.0, 30°C, recombinant mutant H220Q
0.0017
NADPH
-
pH 8.0, 30°C, recombinant mutant H220T
0.0023
NADPH
-
pH 8.0, 30°C, recombinant mutant H220W
0.0024
NADPH
-
pH 8.0, 30°C, recombinant mutant H220Q/K336N
0.011
NADPH
-
pH 8.0, 30°C, recombinant mutant H220Q/K336H
0.011
NADPH
-
pH 8.0, 30°C, recombinant mutant K336H
0.02
NADPH
-
pH 8.0, 30°C, recombinant mutant H220F
0.036
NADPH
-
pH 8.0, 30°C, recombinant mutant H220D
0.17
NADPH
-
pH 8.0, 30°C, recombinant mutant H220E
0.32
NADPH
-
pH 8.0, 30°C, recombinant mutant R217A
0.85
NADPH
-
pH 8.0, 30°C, recombinant mutant R217L
0.04
phenylacetone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
0.059
phenylacetone
-
wild-type enzyme
0.059
phenylacetone
-
30°C, pH 7.5, 0.1 mM NADPH
0.06
phenylacetone
recombinant wild-type enzyme, pH 7.4, 25°C
0.061
phenylacetone
-
25°C, pH 8.5
0.08
phenylacetone
-
wild type enzyme, at pH 7.5 and 37°C
0.17
phenylacetone
recombinant mutant R258A, pH 7.4, 25°C
0.28
phenylacetone
-
25°C, pH 8.5, in the presence of 0.006 mM bovine serum albumin
1.4
phenylacetone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
2.5
phenylacetone
-
mutant P3
3
phenylacetone
-
mutant P1
4
phenylacetone
-
mutant P2
additional information
additional information
-
steady-state kinetics
-
additional information
additional information
Michaelis-Menten kinetics
-
additional information
additional information
steady-state kinetics of wild-type and mutant enzymes
-
additional information
additional information
steady-state kinetics of wild-type and mutant enzymes
-
additional information
additional information
detailed steady-state and pre-steady-state kinetic analysis of the reductive and the oxidative half-reaction of wild-type and mutant enzymes, overview
-
additional information
additional information
-
detailed steady-state and pre-steady-state kinetic analysis of the reductive and the oxidative half-reaction of wild-type and mutant enzymes, overview
-
additional information
additional information
-
steady-state kinetics with NADPH and NADH cofactors, overview
-
additional information
additional information
steady-state kinetic analysis of wild-type and mutant enzymes
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
2
(2-methylphenyl)acetone
-
30°C, pH 7.5, 0.1 mM NADPH
0.023 - 0.23
2-dodecanone
0.07 - 0.5
2-phenylcyclohexanone
0.34
4-hydroxyacetophenone
-
30°C, pH 7.5, 0.1 mM NADPH
0.021 - 1.8
benzylacetone
1.1
bicyclohept-2-en-6-one
-
30°C, pH 7.5, 0.1 mM NADPH
0.0456 - 0.304
cyclohexanone
0.027
Ethionamide
-
25°C, pH 8.5
2.1
methyl 4-tolylsulfide
-
30°C, pH 7.5, 0.1 mM NADPH
additional information
additional information
-
all substrates show a turnover between 1.2 s-1 and 3.6 s-1
-
0.023
2-dodecanone
-
25°C, pH 8.5
0.23
2-dodecanone
-
30°C, pH 7.5, 0.1 mM NADPH
0.067
2-Octanone
recombinant mutant R258M, pH 7.4, 25°C
0.091
2-Octanone
recombinant mutant R258A, pH 7.4, 25°C
0.22
2-Octanone
recombinant wild-type enzyme, pH 7.4, 25°C
1
2-Octanone
-
wild type enzyme, at pH 7.5 and 37°C
2.3
2-Octanone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
2.3
2-Octanone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
0.07
2-phenylcyclohexanone
recombinant wild-type enzyme, pH 7.4, 25°C
0.07
2-phenylcyclohexanone
-
wild type enzyme, at pH 7.5 and 37°C
0.25
2-phenylcyclohexanone
-
mutant P3
0.3
2-phenylcyclohexanone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
0.3
2-phenylcyclohexanone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
0.31
2-phenylcyclohexanone
-
mutant P1
0.5
2-phenylcyclohexanone
-
mutant P2
0.021
benzylacetone
-
25°C, pH 8.5
1.8
benzylacetone
-
30°C, pH 7.5, 0.1 mM NADPH
0.0456
cyclohexanone
mutant I67Y/P440F, pH 8.0, 45°C
0.0653
cyclohexanone
mutant P440F, pH 8.0, 45°C
0.1324
cyclohexanone
mutant I67G/P440F, pH 8.0, 45°C
0.1364
cyclohexanone
mutant I67A/P440F, pH 8.0, 45°C
0.156
cyclohexanone
pH 8.0, 25°C, recombinant mutant S441G/A442P/L443T/S444Q
0.198
cyclohexanone
mutant I67C/P440F, pH 8.0, 45°C
0.304
cyclohexanone
pH 8.0, 25°C, recombinant mutant A442P/L443V
0.9
Cyclopentanone
recombinant wild-type enzyme, pH 7.4, 25°C
0.9
Cyclopentanone
-
wild type enzyme, at pH 7.5 and 37°C
1.6
Cyclopentanone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
1.6
Cyclopentanone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
0.8
NADPH
-
pH 8.0, 30°C, recombinant mutant H220W
0.8
NADPH
-
pH 8.0, 30°C, recombinant mutant R217L
1
NADPH
-
pH 8.0, 30°C, recombinant mutant H220Q/K336N
1.2
NADPH
-
pH 8.0, 30°C, recombinant mutant H220D
1.2
NADPH
-
pH 8.0, 30°C, recombinant mutant H220E
1.2
NADPH
-
pH 8.0, 30°C, recombinant mutant K336H
1.6
NADPH
-
pH 8.0, 30°C, recombinant mutant K336N
1.9
NADPH
-
pH 8.0, 30°C, recombinant mutant H220F
1.9
NADPH
-
pH 8.0, 30°C, recombinant mutant H220Q/K336H
2.2
NADPH
-
pH 8.0, 30°C, recombinant mutant R217A
2.7
NADPH
-
pH 8.0, 30°C, recombinant mutant H220Q
2.9
NADPH
-
pH 8.0, 30°C, recombinant mutant H220T
3.1
NADPH
-
pH 8.0, 30°C, recombinant wild-type PAMO
3.3
NADPH
-
pH 8.0, 30°C, recombinant mutant H220A
3.6
NADPH
-
pH 8.0, 30°C, recombinant mutant H220N
3.8
NADPH
-
pH 8.0, 30°C, recombinant mutant T218A
0.017
phenylacetone
-
25°C, pH 8.5
0.04
phenylacetone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
0.22
phenylacetone
-
mutant P3
0.25
phenylacetone
-
mutant P1
0.26
phenylacetone
-
25°C, pH 8.5, in the presence of 0.006 mM bovine serum albumin
0.4
phenylacetone
-
mutant P2
1.4
phenylacetone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
1.9
phenylacetone
-
wild-type enzyme
1.9
phenylacetone
-
30°C, pH 7.5, 0.1 mM NADPH
2.1
phenylacetone
recombinant mutant R258A, pH 7.4, 25°C
2.4
phenylacetone
recombinant wild-type enzyme, pH 7.4, 25°C
3
phenylacetone
-
wild type enzyme, at pH 7.5 and 37°C
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0.0016 - 0.375
2-phenylcyclohexanone
0.216 - 0.63
cyclohexanone
0.00075 - 1.6
Cyclopentanone
0.043
2-Octanone
recombinant mutant R258A, pH 7.4, 25°C
0.069
2-Octanone
recombinant wild-type enzyme, pH 7.4, 25°C
0.223
2-Octanone
recombinant mutant R258M, pH 7.4, 25°C
0.48
2-Octanone
-
wild type enzyme, at pH 7.5 and 37°C
9.2
2-Octanone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
9.2
2-Octanone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
0.0016
2-phenylcyclohexanone
recombinant wild-type enzyme, pH 7.4, 25°C
0.008
2-phenylcyclohexanone
-
wild type enzyme, at pH 7.5 and 37°C
0.37
2-phenylcyclohexanone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
0.375
2-phenylcyclohexanone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
0.216
cyclohexanone
mutant I67Y/P440F, pH 8.0, 45°C
0.393
cyclohexanone
mutant I67A/P440F, pH 8.0, 45°C
0.436
cyclohexanone
pH 8.0, 25°C, recombinant mutant A442P/L443V
0.449
cyclohexanone
mutant I67C/P440F, pH 8.0, 45°C
0.51
cyclohexanone
mutant I67G/P440F, pH 8.0, 45°C
0.577
cyclohexanone
pH 8.0, 25°C, recombinant mutant S441G/A442P/L443T/S444Q
0.63
cyclohexanone
mutant P440F, pH 8.0, 45°C
0.00075
Cyclopentanone
recombinant wild-type enzyme, pH 7.4, 25°C
0.0016
Cyclopentanone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
0.8
Cyclopentanone
-
wild type enzyme, at pH 7.5 and 37°C
1.6
Cyclopentanone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
1
NADPH
-
pH 8.0, 30°C, recombinant mutant R217L
7
NADPH
-
pH 8.0, 30°C, recombinant mutant H220E
7
NADPH
-
pH 8.0, 30°C, recombinant mutant R217A
33
NADPH
-
pH 8.0, 30°C, recombinant mutant H220D
95
NADPH
-
pH 8.0, 30°C, recombinant mutant H220F
110
NADPH
-
pH 8.0, 30°C, recombinant mutant K336H
170
NADPH
-
pH 8.0, 30°C, recombinant mutant H220Q/K336H
300
NADPH
-
pH 8.0, 30°C, recombinant mutant H220W
420
NADPH
-
pH 8.0, 30°C, recombinant mutant H220Q/K336N
1500
NADPH
-
pH 8.0, 30°C, recombinant mutant K336N
1600
NADPH
-
pH 8.0, 30°C, recombinant mutant H220Q
1700
NADPH
-
pH 8.0, 30°C, recombinant mutant H220T
2700
NADPH
-
pH 8.0, 30°C, recombinant mutant T218A
4000
NADPH
-
pH 8.0, 30°C, recombinant wild-type PAMO
5000
NADPH
-
pH 8.0, 30°C, recombinant mutant H220A
6000
NADPH
-
pH 8.0, 30°C, recombinant mutant H220N
0.029
phenylacetone
recombinant mutant P253F/G254A/R258M/L443F, pH 7.4, 25°C
12.4
phenylacetone
recombinant mutant R258A, pH 7.4, 25°C
35
phenylacetone
-
mutant enzyme P253F/G254A/R258M/L443F, at pH 7.5 and 37°C
37.5
phenylacetone
-
wild type enzyme, at pH 7.5 and 37°C
40
phenylacetone
recombinant wild-type enzyme, pH 7.4, 25°C
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A442G
-
the mutant shows 75% of wild type activity
A442P
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 81% conversion rate
A442P/ L443I/S444Q
random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 43% conversion rate
A442P/ L443V/S444Q
random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 45% conversion rate
A442P/L443I
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 45% conversion rate
A442P/L443L/S444Q
random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 41% conversion rate
A442P/L443T/S444Q
random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 56% conversion rate
A442P/L443V
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 90% conversion rate
A442P/L443W
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 74% conversion rate
A442P/L443W/ S444Q
random and site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 33% conversion rate
C65D/M446I
site-directed mutagenesis
C65D/M446I/Y495I
site-directed mutagenesis, the M446I and Y495I mutations do not have significant influence on the NADPH oxidation activities. The triple mutant, which shows the greatest stability to H2O2, exhibits the highest catalytic activity (kcat) for NADPH oxidation. Thus, the oxidative stability is not markedly related to the NADPH oxidation and H2O2 generation rates
C65D/M446I/Y517I
site-directed mutagenesis, the M446I mutation does not have significant influence on the NADPH oxidation activities. The residual activity of this triple mutant variant remains unchanged during incubation with externally added H2O2. The variant completely loses the catalytic activity after 1 hour when H2O2 is generated in situ from NADPH oxidation. This fast deactivation of the C65D/M446I/Y517I variant leads to oxidation of only 10% of NADPH added. The Y517I mutation in C65D/M446I variant appears to result in blocking of the H2O2 exit and entrance path. The H2O2 generated in the active site might remain there, oxidizing amino acid residues in vicinity of the active site. The low catalytic activity of the C65D/M446I/Y517I variant for NADPH oxidation suggests that the Y517I mutation results in not only blocking of the H2O2 migration path but also modification of the active site structure
C65V
-
the mutant shows 94% of wild type activity
C65V/I67T
-
the mutant shows 47% of wild type activity
C65V/I67T/Q152F/S441A/A442G
-
the mutant shows 31% of wild type activity
C65V/I67T/Q93W
-
the mutant shows 54% of wild type activity
H220A
-
site-directed mutagenesis
H220D
-
site-directed mutagenesis
H220E
-
site-directed mutagenesis, H220E mutant performs worse than wild-type PAMO with both coenzymes NADPH and NADH
H220F
-
site-directed mutagenesis
H220N
-
site-directed mutagenesis, the mutant shows about 3fold improvement in the catalytic efficiency with NADH while the catalytic efficiency with NADPH is hardly affected
H220Q
-
site-directed mutagenesis, the mutant shows about 3fold improvement in the catalytic efficiency with NADH while the catalytic efficiency with NADPH is hardly affected
H220Q/K336H
-
site-directed mutagenesis
H220Q/K336N
-
site-directed mutagenesis
H220T
-
site-directed mutagenesis
H220W
-
site-directed mutagenesis
I339S
-
the mutant shows 81% of wild type activity
I67A
site-directed mutagenesis, the mutant shows no activity with cyclohexanone like the wild-type enzyme
I67A/P440F
site-directed mutagenesis, the mutant shows high activity with cyclohexanone, 95.1% conversion after 4 h
I67C
site-directed mutagenesis, the mutant shows no activity with cyclohexanone like the wild-type enzyme
I67C/P440F
site-directed mutagenesis, the mutant shows high activity with cyclohexanone, 92.4% conversion after 4 h
I67C/P440F/A442F/L443D
site-directed mutagenesis, the mutant shows moderate activity with cyclohexanone
I67C/P440Y
site-directed mutagenesis, the mutant shows moderate activity with cyclohexanone
I67G
site-directed mutagenesis, the mutant shows no activity with cyclohexanone like the wild-type enzyme
I67G/P440F
site-directed mutagenesis, the mutant shows high activity with cyclohexanone, 87.5% conversion after 4 h
I67T
-
the mutant shows 16% of wild type activity
I67T/L338P
-
the mutant shows less than 3% of wild type activity
I67T/L338P/A435Y/A442G
-
the mutant shows less than 3% of wild type activity
I67T/L338P/A435Y/A442G/L443F/S444C
-
the mutant shows less than 3% of wild type activity
I67Y
site-directed mutagenesis, the mutant shows no activity with cyclohexanone like the wild-type enzyme
I67Y/P440F
site-directed mutagenesis, the mutant shows high activity with cyclohexanone, 97.8% conversion after 4 h
I67Y/P440Y
site-directed mutagenesis, the mutant shows high activity with cyclohexanone
K336H
-
site-directed mutagenesis
K336N
-
site-directed mutagenesis
L338P
-
the mutant shows 59% of wild type activity
L443F
-
the mutant shows 65% of wild type activity
L443V
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 53% conversion rate
L443V/S444M
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 53% conversion rate
L443V/S444T
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 57% conversion rate
L447P
-
the mutant shows 85% of wild type activity
P440L
-
higher subtrate variability, temperature optimum at 50C with range from 45-56C
P440N
-
higher subtrate variability, temperature optimum at 50C with range from 45-56C
P440T
-
higher subtrate variability, temperature optimum at 50C with range from 45-56C
P440W
site-directed mutagenesis, the mutant shows high acticity with cyclohexanone
Q152F
-
the mutant shows 35% of wild type activity
Q152F/A442G
-
the mutant shows 37% of wild type activity
Q152F/S441A/A442G
-
the mutant shows 33% of wild type activity
Q93N/P94D/P440F
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at low rate
Q93W
-
the mutant shows 65% of wild type activity
Q93W/A442G/S444C/M446G/L447P
-
the mutant shows 38% of wild type activity
Q93W/S441A/A442G/S444C/M446G/L447P
-
the mutant shows 40% of wild type activity
R217A
-
site-directed mutagenesis
R217L
-
site-directed mutagenesis
R258A
site-directed mutagenesis, when the substrate 2-octanone binds to the R258A mutant, a significant change in the position of the hexyl tail of the substrate is observed, altered substrate specificity compared to wild-type
R258M
site-directed mutagenesis, the R258M mutation significantly affects pose of 2-octanone, since the hexyl tail moves towards M258, substrate specificity compared to wild-type
R337A
site-directed mutagenesis, the mutant is still able to form and stabilize the C4a-peroxyflavin intermediate, but loses the ability to convert phenylacetone or benzyle methylsulfide
R337K
site-directed mutagenesis, the mutant is still able to form and stabilize the C4a-peroxyflavin intermediate, but loses the ability to convert phenylacetone or benzyle methylsulfide
S441A
-
the mutant shows 73% of wild type activity
S441A/A442G
-
the mutant shows 56% of wild type activity
S441A/A442G/S444C/M446G/L447P
-
the mutant shows 41% of wild type activity
S441D/A442E
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 73% conversion rate
S441G/A442P/L443T/S444Q
site-directed mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at about 90% conversion rate
S441G/A442T
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 48% conversion rate
S441H
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 34% conversion rate
S441H/A442P
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 78% conversion rate
S444C
-
the mutant shows 66% of wild type activity
T218A
-
site-directed mutagenesis
V54I
-
the mutant shows 65% of wild type activity
V54I/C65V/I67T/Q93W/I339S/S441A/A442G/S444C/M446G/L447P
-
the mutant shows 5% of wild type activity
V54I/C65V/I67T/Q93W/Q152F/I339S/S441A/A442G/S444C/M446G/L447P
-
the mutant shows less than 3% of wild type activity
V54I/C65V/I67T/Q93W/Q152F/L153G/I339S/S441A/A442G/S444C/M446G/L447P
-
the mutant shows less than 3% of wild type activity
W501A
-
the mutant shows reduced activity compared to the wild type enzyme
A435Y
-
the mutant is active only with bicyclo[3.2.0]hept-2-en-6-one
A435Y
-
the mutant shows less than 3% of wild type activity
C65D
site-directed mutagenesis
C65D
site-directed mutagenesis, the engineered TfPAMO variant acts as an NADPH oxidase, it shows an extremely high rate of uncoupling compared with the wild-type enzyme. The mutant is not effective in stabilizing the C(4alpha)-peroxyflavin intermediate. For TfPAMO C65D, hydrogen peroxide and superoxide levels are highest at pH 9.0
L153G
-
inactive
L153G
-
the mutant shows less than 3% of wild type activity
L443V/S444Q
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 40-45% conversion rate
L443V/S444Q
random/saturation mutagenesis, the mutant is active with cyclohexanone, in contrast to the wild-type enzyme, and catalyzes its conversion to epsilon-caprolactone at 59% conversion rate
M446G
-
enzyme has altered activity and substrate specificity
M446G
-
the mutant retains wild type thermostability and produces an altered substrate binding pocket, leading to substantial changes in substrate specificity and enantioselectivity towards sulfides and ketones
M446G
-
the mutant shows 49% of wild type activity and is able to convert 1-indanone to 1-isochromanone
P253F/G254A/R258M/L443F
-
the mutant shows the same thermostability as the wild type enzyme while it displays an extended substrate spectrum
P253F/G254A/R258M/L443F
site-directed mutagenesis, the engineered mutant quadruple enzyme variant P253F/G254A/R258M/L443F exhibits significantly improved activity towards 2-octanone compared to wild-type. A remarkable movement of L289 is crucial for a reshaping of the active site of the quadruple variant so as to prevent the aliphatic substrate from moving away from the C4a-peroxyflavin, thus enabling it to keep a catalytically relevant pose during the oxygenation process, substrate specificity compared to wild-type
P440F
-
higher subtrate variability, temperature optimum at 50C with range from 45-56C
P440F
site-directed mutagenesis, the mutant shows high activity with cyclohexanone, 50.2% conversion after 4 h
P440H
-
higher subtrate variability, temperature optimum at 50C with range from 45-56C
P440H
site-directed mutagenesis, the mutant shows low activity with cyclohexanone
P440I
-
higher subtrate variability, temperature optimum at 50C with range from 45-56C
P440I
site-directed mutagenesis, the mutant shows low activity with cyclohexanone
P440Y
-
higher subtrate variability, temperature optimum at 50C with range from 45-58C
P440Y
site-directed mutagenesis, the mutant shows low activity with cyclohexanone
additional information
-
construction of three mutants P1-P3 by elimination of a bulge loop region, involving residues Ser441, Ala442, and Leu443, leading to enhanced substrate enantioselectivity of Baeyer-Villiger reactions while maintaining high thermal stability, overview
additional information
-
engineering of three highly stereoselective mutants of the thermally stable phenylacetone monooxygenase as practical catalysts for enantioselective Baeyer-Villiger oxidations of several ketones on a preparative scale under in vitro conditions, optimization of the method including a coupled cofactor-regeneration system, reaction mechanism, overview
additional information
directed evolution of phenylacetone monooxygenase as an active catalyst for the Baeyer-Villiger conversion of cyclohexanone to caprolactone using iterative saturation mutagenesis, mutant screening, overview. Molecular dynamics simulations and induced fit docking of wild-type and mutant enzymes with cyclohexanone. The mutants are used in the whole cell system of Escherichia coli cells
additional information
rational engineering of enzyme PAMO for wide applications in industrial biocatalysis, in particular, in the biotransformation of long-chain aliphatic oils into potential biodiesels
additional information
a rational approach is used to improve the robustness of enzymes, in particular, Baeyer-Villiger monooxygenases (BVMOs) against H2O2. The enzyme access tunnels, which may serve as exit paths for H2O2 from the active site to the bulk, are predicted by using the CAVER and/or protein energy landscape exploration (PELE) software for mutant PAMO_C65D from Thermobifida fusca. The amino acid residues, which are susceptible to oxidation by H2O2 (e.g. methionine and tyrosine) and located in vicinity of the predicted H2O2 migration paths, are substituted with less reactive or inert amino acids (e.g. leucine and isoleucine), leading to design of H2O2-resistant enzyme variants
additional information
addition of the dimerization-docking and anchoring domain (RIDD-RIAD) system to the C-terminus of the NADPH-dependent Baeyer-Villiger monooxygenase phenylacetone monooxygenase (PAMO) and the NADPH-regenerating enzyme (phosphite dehydrogenase, PTDH, EC 1.20.1.1) allowing self-assembly based on specific protein-protein interactions between both peptides and allow tuning of the ratio of the targeted enzymes as the RIAD peptide binds to two RIDD peptides. Several RIDD/RIAD-tagged PAMO and PTDH variants are successfully overproduced in Escherichia coli and subsequently purified. Complementary tagged enzymes are mixed and analyzed for their oligomeric state, stability, and activity. Complexes are formed in the case of some specific combinations (PAMORIAD-PTDHRIDD and PAMORIAD/RIAD-PTDHRIDD). These enzyme complexes display similar catalytic activity when compared with the PTDH-PAMO fusion enzyme. The thermostability of PAMO in these complexes is retained while PTDH displays somewhat lower thermostability
additional information
expanding the substrate scope of a thermostable phenylacetone monooxygenase (PAMO) to cyclohexanone by using site-directed mutagenesis. Several mutants are found to be active with cyclohexanone for which wild-type PAMO shows no activity. There is possible additive or cooperative effect existing between I67 and P440. Based on the thermostable PAMO scaffold, a chimeric PAMO-CHMO enzyme mutant is created, which shows no activity on cyclohexanone
additional information
rational engineering of PAMO for wide applications in industrial biocatalysis, in particular, in the biotransformation of long-chain aliphatic oils into potential biodiesels
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Fraaije, M.W.; Wu, J.; Heuts, D.P.; van Hellemond, E.W.; Spelberg, J.H.; Janssen, D.B.
Discovery of a thermostable Baeyer-Villiger monooxygenase by genome mining
Appl. Microbiol. Biotechnol.
66
393-400
2005
Thermobifida fusca
brenda
Fraaije, M.W.; Kamerbeek, N.M.; Heidekamp, A.J.; Fortin, R.; Janssen, D.B.
The prodrug activator EtaA from Mycobacterium tuberculosis is a Baeyer-Villiger monooxygenase
J. Biol. Chem.
279
3354-3360
2004
Mycobacterium tuberculosis
brenda
Malito, E.; Alfieri, A.; Fraaije, M.W.; Mattevi, A.
Crystal structure of a Baeyer-Villiger monooxygenase
Proc. Natl. Acad. Sci. USA
101
13157-13162
2004
Thermobifida fusca (Q47PU3), Thermobifida fusca
brenda
Bocola, M.; Schulz, F.; Leca, F.; Vogel, A.; Fraaije, M.W.; Reetz, M.T.
Converting phenylacetone monooxygenase into phenylcyclohexanone monooxygenase by rational design: towards practical Baeyer-Villiger monooxygenases
Adv. Synth. Catal.
347
979-986
2005
Thermobifida fusca
-
brenda
Schulz, F.; Leca, F.; Hollmann, F.; Reetz, M.T.
Towards practical biocatalytic Baeyer-Villiger reactions: applying a thermostable enzyme in the gram-scale synthesis of optically active lactones in a two-liquid-phase system
Beilstein J. Org. Chem.
1
10
2005
Thermobifida fusca
brenda
De Gonzalo, G.; Ottolina, G.; Zambianchi, F.; Fraaije, M.W.; Carrea, G.
Biocatalytic properties of Baeyer-Villiger monooxygenases in aqueous-organic media
J. Mol. Catal. B
39
91-97
2006
Thermobifida fusca
-
brenda
de Gonzalo, G.; Torres Pazmino, D.E.; Ottolina, G.; Fraaije, M.W.; Carrea, G.
Oxidations catalyzed by phenylacetone monooxygenase from Thermobifida fusca
Tetrahedron
16
3077-3083
2005
Thermobifida fusca
-
brenda
Rehdorf, J.; Kirschner, A.; Bornscheuer, U.T.
Cloning, expression and characterization of a Baeyer-Villiger monooxygenase from Pseudomonas putida KT2440
Biotechnol. Lett.
29
1393-1398
2007
Pseudomonas putida, Pseudomonas putida KT 2240
brenda
Zambianchi, F.; Fraaije, M.W.; Carrea, G.; de Gonzalo, G.; Rodriguez, C.; Gotor, V.; Ottolina, G.
Titration and assignment of residues that regulate the enantioselectivity of phenylacetone monooxygenase
Adv. Synth. Catal.
349
1327-1331
2007
Thermobifida fusca (Q47PU3)
-
brenda
Torres Pazmino, D.E.; Baas, B.J.; Janssen, D.B.; Fraaije, M.W.
Kinetic mechanism of phenylacetone monooxygenase from Thermobifida fusca
Biochemistry
47
4082-4093
2008
Thermobifida fusca (Q47PU3), Thermobifida fusca
brenda
Rodriguez, C.; de Gonzalo, G.; Torres Pazmino, D.E.; Fraaije, M.W.; Gotor, V.
Selective Baeyer-Villiger oxidation of racemic ketones in aqueous-organic media catalyzed by phenylacetone monooxygenase
Tetrahedron Asymmetry
19
197-203
2008
Thermobifida fusca
-
brenda
Rioz-Martinez, A.; De Gonzal, D.G.; Torres Pazmino, D.; Fraaije, M.; Gotor, V.
Enzymatic Baeyer-Villiger oxidation of benzo-fused ketones: Formation of regiocomplementary lactones
Eur. J. Org. Chem.
2009
2526-2532
2009
Pseudomonas fluorescens, Thermobifida fusca
-
brenda
Reetz, M.T.; Wu, S.
Laboratory evolution of robust and enantioselective Baeyer-Villiger monooxygenases for asymmetric catalysis
J. Am. Chem. Soc.
131
15424-15432
2009
Thermobifida fusca
brenda
Dudek, H.M.; Torres Pazmino, D.E.; Rodriguez, C.; de Gonzalo, G.; Gotor, V.; Fraaije, M.W.
Investigating the coenzyme specificity of phenylacetone monooxygenase from Thermobifida fusca
Appl. Microbiol. Biotechnol.
88
1135-1143
2010
Thermobifida fusca
brenda
Dudek, H.M.; de Gonzalo, G.; Pazmino, D.E.; Stepniak, P.; Wyrwicz, L.S.; Rychlewski, L.; Fraaije, M.W.
Mapping the substrate binding site of phenylacetone monooxygenase from Thermobifida fusca by mutational analysis
Appl. Environ. Microbiol.
77
5730-5738
2011
Thermobifida fusca
brenda
Dudek, H.M.; Fink, M.J.; Shivange, A.V.; Dennig, A.; Mihovilovic, M.D.; Schwaneberg, U.; Fraaije, M.W.
Extending the substrate scope of a Baeyer-Villiger monooxygenase by multiple-site mutagenesis
Appl. Microbiol. Biotechnol.
98
4009-4020
2013
Thermobifida fusca
brenda
de Gonzalo, G.; Rodriguez, C.; Rioz-Martinez, A.; Gotor, V.
Improvement of the biocatalytic properties of one phenylacetone monooxygenase mutant in hydrophilic organic solvents
Enzyme Microb. Technol.
50
43-49
2012
Thermobifida fusca
brenda
Andrade, L.; Pedrozo, E.; Leite, H.; Brondani, P.
Oxidation of organoselenium compounds. A study of chemoselectivity of phenylacetone monooxygenase
J. Mol. Catal. B
73
63-66
2011
Thermobifida fusca
-
brenda
Rodriguez, C.; De Gonzalo, G.; Gotor, V.
Optimization of oxidative bioconversions catalyzed by phenylacetone monooxygenase from Thermobifida fusca
J. Mol. Catal. B
74
138-143
2012
Thermobifida fusca
-
brenda
Parra, L.P.; Acevedo, J.P.; Reetz, M.T.
Directed evolution of phenylacetone monooxygenase as an active catalyst for the Baeyer-Villiger conversion of cyclohexanone to caprolactone
Biotechnol. Bioeng.
112
1354-1364
2015
Thermobifida fusca (Q47PU3)
brenda
Carvalho, A.T.P.; Dourado, D.F.A.R.; Skvortsov, T.; de Abreu, M.; Ferguson, L.J.; Quinn, D.J.; Moody, T.S.; Huang, M.
Catalytic mechanism of phenylacetone monooxygenases for non-native linear substrates
Phys. Chem. Chem. Phys.
19
26851-26861
2017
Thermobifida fusca (Q47PU3)
brenda
Gran-Scheuch, A.; Parra, L.; Fraaije, M.
Systematic assessment of uncoupling in flavoprotein oxidases and monooxygenases
ACS Sust. Chem. Eng.
FEHLT
0000
2021
Thermobifida fusca (Q47PU3)
-
brenda
Seo, E.; Kim, M.; Park, S.; Park, S.; Oh, D.; Bornscheuer, U.; Park, J.
Enzyme access tunnel engineering in Baeyer-Villiger monooxygenases to improve oxidative stability and biocatalyst performance
Adv. Synth. Catal.
364
555-564
2022
Thermobifida fusca (Q47PU3)
-
brenda
Parshin, P.D.; Pometun, A.A.; Martysuk, U.A.; Kleymenov, S.Y.; Atroshenko, D.L.; Pometun, E.V.; Savin, S.S.; Tishkov, V.I.
Effect of His6-tag position on the expression and properties of phenylacetone monooxygenase from Thermobifida fusca
Biochemistry (Moscow)
85
575-582
2020
Thermobifida fusca (Q47PU3), Thermobifida fusca
brenda
Purwani, N.N.; Martin, C.; Savino, S.; Fraaije, M.W.
Modular assembly of phosphite dehydrogenase and phenylacetone monooxygenase for tuning cofactor regeneration
Biomolecules
11
905
2021
Thermobifida fusca (Q47PU3)
brenda
Yang, G.; Cang, R.; Shen, L.; Xue, F.; Huang, H.; Zhang, Z.
Expanding the substrate scope of phenylacetone monooxygenase from Thermobifida fusca towards cyclohexanone by protein engineering
Catal. Commun.
119
159-163
2019
Thermobifida fusca (Q47PU3)
-
brenda
Carvalho, A.T.P.; Dourado, D.F.A.R.; Skvortsov, T.; de Abreu, M.; Ferguson, L.J.; Quinn, D.J.; Moody, T.S.; Huang, M.
Spatial requirement for PAMO for transformation of non-native linear substrates
Phys. Chem. Chem. Phys.
20
2558-2570
2018
Thermobifida fusca (Q47PU3)
brenda