1.14.13.92 (2-methylphenyl)acetone + NADPH + O2 - Thermobifida fusca ? + NADP+ + O2 - ? 375417 1.14.13.92 (2R,3S)-3-methyl-2-pentylcyclopentanone + NADPH + H+ + O2 less than 5% conversion Thermobifida fusca ? - ? 427751 1.14.13.92 (R)-1-acetoxy-phenylacetone + NADPH + O2 - Thermobifida fusca (R)-1-hydroxy-1-phenylacetone + NADP+ + H2O - ? 386880 1.14.13.92 (R)-2-acetoxyphenylacetonitrile + NADPH + H+ + O2 enantioselective reaction Thermobifida fusca ? - ? 395127 1.14.13.92 (R)-3-(4-bromophenyl)butan-2-one + NADPH + H+ + O2 enantioselective reaction Thermobifida fusca ? - ? 395130 1.14.13.92 (S)-1-(3-trifluoromethylphenyl)ethyl acetate + NADPH + H+ + O2 enantioselective reaction Thermobifida fusca ? - ? 395154 1.14.13.92 (S)-nicotine + NADPH + O2 - Thermobifida fusca ? - ? 386912 1.14.13.92 1-bromo-indanone + NADPH + O2 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 Pseudomonas fluorescens 6-bromoisochroman-1-one + NADP+ + H2O - ? 403215 1.14.13.92 1-indanone + NADPH + H+ + O2 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 Pseudomonas fluorescens 3,4-dihydrocoumarin + NADP+ + H2O - ? 403247 1.14.13.92 1-indanone + NADPH + H+ + O2 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 Thermobifida fusca 1-isochromanone + NADP+ + H2O - ? 403249 1.14.13.92 1-tetralone + NADPH + O2 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 Pseudomonas fluorescens 4,5-dihydro-1-benzoxepin-2(3H)-one + NADP+ + H2O - ? 403296 1.14.13.92 1-[3-(benzylselanyl)phenyl]ethanone + NADPH + H+ + O2 more than 99% conversion Thermobifida fusca 1-(3-(benzylseleninyl)phenyl)ethanone + NADP+ + H2O - ? 427839 1.14.13.92 1-[3-(methylselanyl)phenyl]ethanone + NADPH + H+ + O2 76% conversion Thermobifida fusca 1-(3-(methylseleninyl)phenyl)ethanone + NADP+ + H2O - ? 427840 1.14.13.92 1-[4-(benzylselanyl)phenyl]ethanone + NADPH + H+ + O2 more than 99% conversion Thermobifida fusca 1-(4-(benzylseleninyl)phenyl)ethanone + NADP+ + H2O - ? 427841 1.14.13.92 1-[4-(methylselanyl)phenyl]ethanone + NADPH + H+ + O2 more than 99% conversion Thermobifida fusca 1-(4-(methylseleninyl)phenyl)ethanone + NADP+ + H2O - ? 427842 1.14.13.92 2-benzylcyclopentanone + NADPH + H+ + O2 about 10% conversion Thermobifida fusca ? - ? 427895 1.14.13.92 2-decanone + NADPH + O2 - Mycobacterium tuberculosis methyl nonanoate + octyl acetate + NADP+ + H2O - ? 375664 1.14.13.92 2-decanone + NADPH + O2 - Pseudomonas putida ? + NADP+ + H2O - ? 390128 1.14.13.92 2-decanone + NADPH + O2 - Pseudomonas putida KT 2240 ? + NADP+ + H2O - ? 390128 1.14.13.92 2-dodecanone + NADPH + O2 - Thermobifida fusca ? + NADP+ - ? 375666 1.14.13.92 2-dodecanone + NADPH + O2 - Mycobacterium tuberculosis nonyl acetate + methyl decanoate + NADP+ + H2O - ? 375667 1.14.13.92 2-dodecanone + NADPH + O2 - Pseudomonas putida ? + NADP+ + H2O - ? 390129 1.14.13.92 2-dodecanone + NADPH + O2 - Pseudomonas putida KT 2240 ? + NADP+ + H2O - ? 390129 1.14.13.92 2-heptanone + NADPH + O2 - Mycobacterium tuberculosis pentyl acetate + NADP+ + H2O - ? 375669 1.14.13.92 2-hexanone + NADPH + O2 - Mycobacterium tuberculosis butyl acetate + NADP+ + H2O - ? 375670 1.14.13.92 2-indanone + NADPH + H+ + O2 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 Thermobifida fusca 3-isochromanone + NADP+ + H2O - ? 403560 1.14.13.92 2-methylcyclohexanone + NADPH + H+ + O2 no activity with wild-type PAMO, but low to increased activity with enzyme mutants Thermobifida fusca ? + NADP+ + H2O - ? 461983 1.14.13.92 2-methylphenylcyclohexanone + NADPH + O2 mutant P3 prefers the R-isomer Thermobifida fusca 7-benzyloxepan-2-one + NADP+ + H2O - ? 387187 1.14.13.92 2-nonanone + NADPH + H+ + O2 less than 40% conversion Thermobifida fusca ? - ? 427929 1.14.13.92 2-nonanone + NADPH + O2 - Pseudomonas putida ? + NADP+ + H2O - ? 390131 1.14.13.92 2-nonanone + NADPH + O2 - Pseudomonas putida KT 2240 ? + NADP+ + H2O - ? 390131 1.14.13.92 2-octanone + NADPH + H+ + O2 - Thermobifida fusca ? - ? 427936 1.14.13.92 2-octanone + NADPH + H+ + O2 - Thermobifida fusca ? + NADP+ + H2O - ? 442816 1.14.13.92 2-octanone + NADPH + H+ + O2 - Thermobifida fusca heptylacetate + NADP+ + H2O - ? 461992 1.14.13.92 2-octanone + NADPH + O2 - Mycobacterium tuberculosis heptyl acetate + NADP+ + H2O - ? 375698 1.14.13.92 2-phenylcyclohexanone + NADPH + H+ + O2 - Thermobifida fusca ? - ? 427945 1.14.13.92 2-phenylcyclohexanone + NADPH + H+ + O2 very low activity with wild-type PAMO, significantly increased activity with enzyme mutant P253F/G254A/R258M/L443F Thermobifida fusca ? + NADP+ + H2O - ? 461999 1.14.13.92 2-phenylcyclohexanone + NADPH + O2 molecular modeling of the Criegee intermediate, the wild-type enzyme prefers the S-isomer, while mutants P1-P3 all prefer the R-isomer Thermobifida fusca 7-phenyloxepan-2-one + NADP+ + H2O - ? 387224 1.14.13.92 2-phenylpropionaldehyde + NADPH + H+ + O2 - Thermobifida fusca ? - ? 395625 1.14.13.92 2-undecanone + NADPH + O2 - Pseudomonas putida ? + NADP+ + H2O - ? 390136 1.14.13.92 3-(3-trifluoromethylphenyl)butan-2-one + NADPH + H+ + O2 enantioselective reaction Thermobifida fusca ? - ? 395694 1.14.13.92 3-(4-chlorophenyl)cyclobutanone + NADPH + H+ + O2 about 40% conversion Thermobifida fusca ? - ? 427968 1.14.13.92 3-benzylcyclobutanone + NADPH + H+ + O2 about 45% conversion Thermobifida fusca ? - ? 427969 1.14.13.92 3-decanone + NADPH + O2 - Pseudomonas putida ? + NADP+ + H2O - ? 390140 1.14.13.92 3-octanone + NADPH + H+ + O2 - Thermobifida fusca ? - ? 427993 1.14.13.92 3-octanone + NADPH + O2 - Mycobacterium tuberculosis ethyl hexanoate + pentyl propanoate + NADP+ + H2O - ? 375832 1.14.13.92 3-phenyl-2-butanone + NADPH + H+ + O2 enantioselective reaction Thermobifida fusca (R)-3-phenylbutan-2-one + (S)-1-phenyethyl acetate - ? 398241 1.14.13.92 3-phenylcyclobutanone + NADPH + H+ + O2 about 70% conversion Thermobifida fusca ? - ? 427997 1.14.13.92 3-phenylpenta-2,4-dione + NADPH + O2 - Thermobifida fusca (R)-phenylacetylcarbinol + NADP+ + H2O the product is a well-known precursor in the synthesis of ephedrine and pseudoephedrine ? 387396 1.14.13.92 4-decanone + NADPH + O2 - Pseudomonas putida ? + NADP+ + H2O - ? 390146 1.14.13.92 4-decanone + NADPH + O2 best substrate Pseudomonas putida ? + NADP+ + H2O - ? 390146 1.14.13.92 4-heptanone + NADPH + O2 - Mycobacterium tuberculosis propanyl butanoate + NADP+ + H2O - ? 375881 1.14.13.92 4-hydroxyacetophenone + NADPH + O2 - Thermobifida fusca acetic acid 4-hydroxyphenyl ester + NADP+ + O2 - ? 375885 1.14.13.92 4-methylcyclohexanone + NADPH + H+ + O2 no activity with wild-type PAMO, but low to increased activity with enzyme mutants Thermobifida fusca ? + NADP+ + H2O - ? 462088 1.14.13.92 4-phenylcyclohexanone + NADPH + O2 - Thermobifida fusca 4-phenyl-hexano-6-lactone + NADP+ + H2O - ? 387542 1.14.13.92 6-methoxy-1-indanone + NADPH + O2 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 Pseudomonas fluorescens ? + NADP+ + H2O - ? 404067 1.14.13.92 alpha-acetylphenylacetonitrile + NADPH + H+ + O2 enantioselective reaction Thermobifida fusca (R)-2-acetoxyphenylacetonitrile + NADP+ + H2O enantiopure product formation ? 396382 1.14.13.92 benzocyclobutanone + NADPH + O2 reaction is performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH Thermobifida fusca 2-coumaranone + NADP+ + H2O - ? 404464 1.14.13.92 benzocyclobutanone + NADPH + O2 reaction was performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH Pseudomonas fluorescens 2-coumaranone + NADP+ + H2O - ? 404464 1.14.13.92 benzylacetone + NADPH + O2 - Mycobacterium tuberculosis ? + NADP+ + H2O - ? 376381 1.14.13.92 benzylacetone + NADPH + O2 - Thermobifida fusca ? + NADP+ + H2O - ? 376381 1.14.13.92 benzylacetone + NADPH + O2 low activity Thermobifida fusca ? - ? 388193 1.14.13.92 bicyclohept-2-en-6-one + NADPH + O2 - Mycobacterium tuberculosis 3-oxabicyclo[3.3.0]oct-6-en-2-one + NADP+ + H2O - ? 376413 1.14.13.92 bicyclohept-2-en-6-one + NADPH + O2 - Thermobifida fusca ? + NADP+ + O2 - ? 376414 1.14.13.92 bicyclo[2.2.1]heptan-2-one + NADPH + H+ + O2 about 5% conversion Thermobifida fusca ? - ? 428444 1.14.13.92 bicyclo[3.2.0]hept-2-en-6-one + NADPH + H+ + O2 - Thermobifida fusca ? - ? 428445 1.14.13.92 cycloheptanone + NADPH + H+ + O2 no activity with wild-type PAMO, but low to increased activity with enzyme mutants Thermobifida fusca ? + NADP+ + H2O - ? 462389 1.14.13.92 cyclohexanone + NADPH + H+ + O2 substrate of enzyme mutants, not of wild-type, overview Thermobifida fusca epsilon-caprolactone + NADP+ + H2O - ? 394811 1.14.13.92 cyclohexanone + NADPH + H+ + O2 no activity with wild-type PAMO, but low to increased activity with enzyme mutants Thermobifida fusca epsilon-caprolactone + NADP+ + H2O - ? 394811 1.14.13.92 cyclohexanone + NADPH + H+ + O2 - Thermobifida fusca ? - ? 396809 1.14.13.92 cyclopentanone + NADPH + H+ + O2 - Thermobifida fusca 5-valerolactone + NADP+ + H2O - ? 376586 1.14.13.92 cyclopentanone + NADPH + H+ + O2 very low activity with wild-type PAMO, significantly increased activity with enzyme mutant P253F/G254A/R258M/L443F Thermobifida fusca 5-valerolactone + NADP+ + H2O - ? 376586 1.14.13.92 cyclopentanone + NADPH + H+ + O2 - Thermobifida fusca ? - ? 396813 1.14.13.92 diketone + NADPH + O2 - Thermobifida fusca (R)-1-acetoxy-phenylacetone + NADP+ + H2O - ? 388559 1.14.13.92 ethionamide + NADPH + O2 - Mycobacterium tuberculosis ? + NADP+ + H2O - ? 376767 1.14.13.92 ethionamide + NADPH + O2 - Thermobifida fusca ? + NADP+ + O2 - ? 376768 1.14.13.92 methyl 4-tolylsulfide + NADPH + O2 - Thermobifida fusca ? + NADP+ + O2 - ? 377158 1.14.13.92 methyl-p-tolylsulfide + NADPH + O2 - Mycobacterium tuberculosis ? + NADP+ + H2O - ? 377167 1.14.13.92 additional information substrate specificity of wild-type and mutant enzymes, overview Thermobifida fusca ? - - 89 1.14.13.92 additional information enzyme activity in a variety of different aqueous–organic media using organic sulfides as substrates, enantioselectivity, overview Thermobifida fusca ? - ? 89 1.14.13.92 additional information substrate selectivity and stereospecificity of wild-type and mutant enzymes, overview Thermobifida fusca ? - ? 89 1.14.13.92 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 Thermobifida fusca ? - ? 89 1.14.13.92 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 Pseudomonas putida ? - ? 89 1.14.13.92 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 Pseudomonas putida ? - ? 89 1.14.13.92 additional information PAMO is an FAD-containing Baeyer-Villiger monooxygenase Thermobifida fusca ? - ? 89 1.14.13.92 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 Thermobifida fusca ? - ? 89 1.14.13.92 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 Thermobifida fusca ? - ? 89 1.14.13.92 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 Thermobifida fusca ? - - 89 1.14.13.92 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 Thermobifida fusca ? - - 89 1.14.13.92 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 Thermobifida fusca ? - - 89 1.14.13.92 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 Pseudomonas putida KT 2240 ? - ? 89 1.14.13.92 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 Pseudomonas putida KT 2240 ? - ? 89 1.14.13.92 N,N-dimethylbenzylamine + NADPH + O2 - Thermobifida fusca N,N-dimethylbenzylamine N-oxide + NADP+ + H2O - ? 389200 1.14.13.92 phenylacetone + NADPH + H+ + O2 - Mycobacterium tuberculosis benzyl acetate + NADP+ + H2O - ? 377503 1.14.13.92 phenylacetone + NADPH + H+ + O2 - Thermobifida fusca benzyl acetate + NADP+ + H2O - ? 377503 1.14.13.92 phenylacetone + NADPH + H+ + O2 enantioselective reaction, regulation mechanism, overview Thermobifida fusca benzyl acetate + NADP+ + H2O - ? 377503 1.14.13.92 phenylacetone + NADPH + O2 reaction is performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH Thermobifida fusca benzyl acetate + NADP+ + H2O - ? 406383 1.14.13.92 phenylacetone + NADPH + O2 reaction is performed in presence of 2 U secondary alcohol dehydrogenase from Thermoanaerobacter ethanolicus and isopropanol to recover NADPH Thermobifida fusca benzyl acetate + NADP+ + H2O - ? 406383 1.14.13.92 phenylacetone + NADPH + O2 reaction was performed in presence of 10 U glucose-6-phosphate dehydrogenase and glucose-6-phosphate to recover NADPH Pseudomonas fluorescens benzyl acetate + NADP+ + H2O - ? 406383 1.14.13.92 phenylboronic acid + NADPH + O2 formation of phenol Thermobifida fusca ? - ? 389544 1.14.13.92 rac-2-ethylcyclohexanone + NADPH + O2 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 Thermobifida fusca benzyl acetate + NADP+ + H2O - ? 406548 1.14.13.92 rac-3-methyl-4-phenylbutan-2-one + NADH + H+ + O2 enantioselective reaction by PAMO Thermobifida fusca (2R)-1-phenylpropan-2-yl acetate + NAD+ + H2O - ? 415177 1.14.13.92 rac-3-methyl-4-phenylbutan-2-one + NADPH + H+ + O2 enantioselective reaction by PAMO Thermobifida fusca (2R)-1-phenylpropan-2-yl acetate + NADP+ + H2O - ? 415178 1.14.13.92 rac-bicyclo [3.2.0]hept-2-en-6-one + NADPH + O2 activity and stereoselectivity of wild-type and mutant enzymes, overview Thermobifida fusca ? - ? 389642 1.14.13.92 thioanisole + NADH + H+ + O2 low activity, less enantioselective reaction Thermobifida fusca thioanisole sulfoxide + NAD+ + H2O - ? 415373 1.14.13.92 thioanisole + NADPH + H+ + O2 - Thermobifida fusca methyl phenyl sulfoxide + NADP+ + H2O - ? 389879 1.14.13.92 thioanisole + NADPH + H+ + O2 - Thermobifida fusca ? - ? 398417 1.14.13.92 thioanisole + NADPH + H+ + O2 the asymmetric oxidation of thioanisole to sulfoxide is accompanied by the overoxidation to achiral sulfone Thermobifida fusca ? - ? 398417 1.14.13.92 thioanisole + NADPH + H+ + O2 enantioselective reaction Thermobifida fusca thioanisole sulfoxide + NADP+ + H2O mainly (R)-sulfoxide ? 415374