HAPMO_PSEFL
ID HAPMO_PSEFL Reviewed; 640 AA.
AC Q93TJ5;
DT 15-MAY-2007, integrated into UniProtKB/Swiss-Prot.
DT 01-DEC-2001, sequence version 1.
DT 03-AUG-2022, entry version 82.
DE RecName: Full=4-hydroxyacetophenone monooxygenase;
DE Short=HAPMO;
DE EC=1.14.13.84;
DE AltName: Full=Baeyer-Villiger monooxygenase;
DE Short=BVMO;
GN Name=hapE;
OS Pseudomonas fluorescens.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=294;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-21, FUNCTION,
RP MUTAGENESIS OF GLY-490, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, AND
RP SUBUNIT.
RC STRAIN=ACB;
RX PubMed=11322873; DOI=10.1046/j.1432-1327.2001.02137.x;
RA Kamerbeek N.M., Moonen M.J.H., van der Ven J.G.M., van Berkel W.J.H.,
RA Fraaije M.W., Janssen D.B.;
RT "4-hydroxyacetophenone monooxygenase from Pseudomonas fluorescens ACB. A
RT novel flavoprotein catalyzing Baeyer-Villiger oxidation of aromatic
RT compounds.";
RL Eur. J. Biochem. 268:2547-2557(2001).
RN [2]
RP MUTAGENESIS OF HIS-296 AND TRP-300.
RC STRAIN=ACB;
RX PubMed=11997015; DOI=10.1016/s0014-5793(02)02623-6;
RA Fraaije M.W., Kamerbeek N.M., van Berkel W.J.H., Janssen D.B.;
RT "Identification of a Baeyer-Villiger monooxygenase sequence motif.";
RL FEBS Lett. 518:43-47(2002).
RN [3]
RP FUNCTION, SUBSTRATE SPECIFICITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=ACB;
RX PubMed=12514023; DOI=10.1128/aem.69.1.419-426.2003;
RA Kamerbeek N.M., Olsthoorn A.J.J., Fraaije M.W., Janssen D.B.;
RT "Substrate specificity and enantioselectivity of 4-hydroxyacetophenone
RT monooxygenase.";
RL Appl. Environ. Microbiol. 69:419-426(2003).
RN [4]
RP MUTAGENESIS OF ARG-339; LYS-439 AND ARG-440, KINETIC PARAMETERS, AND
RP ACTIVITY REGULATION.
RC STRAIN=ACB;
RX PubMed=15153101; DOI=10.1111/j.1432-1033.2004.04126.x;
RA Kamerbeek N.M., Fraaije M.W., Janssen D.B.;
RT "Identifying determinants of NADPH specificity in Baeyer-Villiger
RT monooxygenases.";
RL Eur. J. Biochem. 271:2107-2116(2004).
RN [5]
RP MASS SPECTROMETRY, AND REACTION MECHANISM.
RX PubMed=16049018; DOI=10.1074/jbc.m503758200;
RA van den Heuvel R.H.H., Tahallah N., Kamerbeek N.M., Fraaije M.W.,
RA van Berkel W.J.H., Janssen D.B., Heck A.J.R.;
RT "Coenzyme binding during catalysis is beneficial for the stability of 4-
RT hydroxyacetophenone monooxygenase.";
RL J. Biol. Chem. 280:32115-32121(2005).
CC -!- FUNCTION: Catalyzes a Baeyer-Villiger oxidation reaction, i.e. the
CC insertion of an oxygen atom into a carbon-carbon bond adjacent to a
CC carbonyl, which converts ketones to esters. Can oxidize a wide range of
CC acetophenone derivatives. Highest activity occurs with compounds
CC bearing an electron-donating substituent at the para position of the
CC aromatic ring, e.g. 4-hydroxyacetophenone and 4-aminoacetophenone,
CC leading to the formation of 4-hydroxyphenyl acetate and 4-aminophenyl
CC acetate, respectively. Is also able to oxidize sulfides.
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4'-hydroxyacetophenone + H(+) + NADPH + O2 = 4-acetoxyphenol +
CC H2O + NADP(+); Xref=Rhea:RHEA:22916, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:28032,
CC ChEBI:CHEBI:31128, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349;
CC EC=1.14.13.84;
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:11322873};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:11322873};
CC -!- ACTIVITY REGULATION: Inhibited by amino-NADP(+).
CC {ECO:0000269|PubMed:15153101}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=12 uM for NADPH (at pH 7.5) {ECO:0000269|PubMed:11322873,
CC ECO:0000269|PubMed:12514023, ECO:0000269|PubMed:15153101};
CC KM=1.44 mM for NADH (at pH 7.5) {ECO:0000269|PubMed:11322873,
CC ECO:0000269|PubMed:12514023, ECO:0000269|PubMed:15153101};
CC KM=9.2 uM for 4-hydroxyacetophenone (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=2.4 uM for 4-hydroxypropiophenone (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=101 uM for 4-hydroxybenzaldehyde (at pH 8.0)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=0.82 uM for 4-aminoacetophenone (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=1.04 mM for 4-fluoroacetophenone (at pH 8.0)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=161 uM for 4-methylacetophenone (at pH 8.0)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=541 uM for 4-methoxyacetophenone (at pH 8.0)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=610 uM for 2-hydroxyacetophenone (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=1.4 mM for 3-hydroxyacetophenone (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=1.6 mM for benzaldehyde (at pH 7.5) {ECO:0000269|PubMed:11322873,
CC ECO:0000269|PubMed:12514023, ECO:0000269|PubMed:15153101};
CC KM=2.27 mM for acetophenone (at pH 8.0) {ECO:0000269|PubMed:11322873,
CC ECO:0000269|PubMed:12514023, ECO:0000269|PubMed:15153101};
CC KM=530 uM for propiophenone (at pH 7.5) {ECO:0000269|PubMed:11322873,
CC ECO:0000269|PubMed:12514023, ECO:0000269|PubMed:15153101};
CC KM=2 mM for butyrophenone (at pH 7.5) {ECO:0000269|PubMed:11322873,
CC ECO:0000269|PubMed:12514023, ECO:0000269|PubMed:15153101};
CC KM=540 uM for isobutyrophenone (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=1.4 mM for methylphenyl sulfide (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=370 uM for methyl 4-tolyl sulfide (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=1.2 mM for 2-acetylpyridine (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=1.9 mM for 4-acetylpyridine (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=330 uM for 2-acetylpyrrole (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=410 uM for 2-pyrrole carboxaldehyde (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=4.8 mM for acetylcyclohexane (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=3 mM for cyclohexane carboxaldehyde (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=29 mM for hydroxyacetone (at pH 7.5) {ECO:0000269|PubMed:11322873,
CC ECO:0000269|PubMed:12514023, ECO:0000269|PubMed:15153101};
CC KM=23 mM for 3-chloro-2-butanone (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=4.9 mM for 2,4-pentanedione (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=3.3 mM for rac-bicyclo[3.2.0]hept-2-en-6-one (at pH 7.5)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC KM=380 uM for 4-hydroxy-3-methylacetophenone (at pH 8.0)
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023,
CC ECO:0000269|PubMed:15153101};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:11322873,
CC ECO:0000269|PubMed:12514023};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius.
CC {ECO:0000269|PubMed:11322873, ECO:0000269|PubMed:12514023};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:11322873}.
CC -!- MASS SPECTROMETRY: Mass=77610; Mass_error=10; Method=Electrospray;
CC Note=The measured mass is that of a monomer in complex with FAD
CC cofactor.; Evidence={ECO:0000269|PubMed:16049018};
CC -!- SIMILARITY: Belongs to the FAD-binding monooxygenase family.
CC {ECO:0000305}.
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DR EMBL; AF355751; AAK54073.1; -; Genomic_DNA.
DR AlphaFoldDB; Q93TJ5; -.
DR SMR; Q93TJ5; -.
DR KEGG; ag:AAK54073; -.
DR BioCyc; MetaCyc:MON-17194; -.
DR BRENDA; 1.14.13.84; 5121.
DR SABIO-RK; Q93TJ5; -.
DR GO; GO:0033767; F:4-hydroxyacetophenone monooxygenase activity; IEA:UniProtKB-EC.
DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IEA:InterPro.
DR GO; GO:0004499; F:N,N-dimethylaniline monooxygenase activity; IEA:InterPro.
DR GO; GO:0050661; F:NADP binding; IEA:InterPro.
DR Gene3D; 3.50.50.60; -; 2.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR020946; Flavin_mOase-like.
DR Pfam; PF00743; FMO-like; 1.
DR SUPFAM; SSF51905; SSF51905; 2.
PE 1: Evidence at protein level;
KW Direct protein sequencing; FAD; Flavoprotein; Monooxygenase; NADP;
KW Oxidoreductase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:11322873"
FT CHAIN 2..640
FT /note="4-hydroxyacetophenone monooxygenase"
FT /id="PRO_0000287886"
FT BINDING 152
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250"
FT BINDING 171
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250"
FT BINDING 180
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250"
FT BINDING 191..192
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250"
FT BINDING 197
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250"
FT BINDING 239
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250"
FT BINDING 275
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250"
FT SITE 440
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000255"
FT MUTAGEN 296
FT /note="H->A: Almost no activity."
FT /evidence="ECO:0000269|PubMed:11997015"
FT MUTAGEN 300
FT /note="W->A,Y: Impaired folding."
FT /evidence="ECO:0000269|PubMed:11997015"
FT MUTAGEN 339
FT /note="R->A: Largely decreased affinity for NADPH."
FT /evidence="ECO:0000269|PubMed:15153101"
FT MUTAGEN 439
FT /note="K->A,F: 100-fold decrease in catalytic efficiency
FT with NADPH. 4-fold higher efficiency with NADH."
FT /evidence="ECO:0000269|PubMed:15153101"
FT MUTAGEN 439
FT /note="K->N: 20-fold decrease in catalytic efficiency with
FT NADPH. 6-fold higher efficiency with NADH."
FT /evidence="ECO:0000269|PubMed:15153101"
FT MUTAGEN 439
FT /note="K->P: 20-fold decrease in catalytic activity."
FT /evidence="ECO:0000269|PubMed:15153101"
FT MUTAGEN 440
FT /note="R->A: No activity. Retains high affinity toward
FT NADPH."
FT /evidence="ECO:0000269|PubMed:15153101"
FT MUTAGEN 490
FT /note="G->A: Retains activity, but shows impaired binding
FT properties of NADPH."
FT /evidence="ECO:0000269|PubMed:11322873"
SQ SEQUENCE 640 AA; 71957 MW; 60750A318723DA6A CRC64;
MSAFNTTLPS LDYDDDTLRE HLQGADIPTL LLTVAHLTGD LQILKPNWKP SIAMGVARSG
MDLETEAQVR EFCLQRLIDF RDSGQPAPGR PTSDQLHILG TWLMGPVIEP YLPLIAEEAV
TAEEDLRAPR WHKDHVASGR DFKVVIIGAG ESGMIAALRF KQAGVPFVIY EKGNDVGGTW
RENTYPGCRV DINSFWYSFS FARGIWDDCF APAPQVFAYM QAVAREHGLY EHIRFNTEVS
DAHWDESTQR WQLLYRDSEG QTQVDSNVVV FAVGQLNRPM IPAIPGIETF KGPMFHSAQW
DHDVDWSGKR VGVIGTGASA TQFIPQLAQT AAELKVFART TNWLLPTPDL HEKISDSCKW
LLAHVPHYSL WYRVAMAMPQ SVGFLEDVMV DVGYPPTELA VSARNDRLRQ DISAWMEPQF
ADRPDLREVL IPDSPVGGKR IVRDNGTWIS TLKRDNVSMI RQPIEVITPK GICCVDGTEH
EFDLIVYGTG FHASKFLMPI NVTGRDGVAL HDVWKGDDAR AYLGMTVPQF PNMFCMYGPN
TGLVVYSTVI QFSEMTASYI VDAVRLLLEG GHQSMEVKTP VFESYNQRVD EGNALRAWGF
SKVNSWYKNS KGRVTQNFPF TAVEFWQRTH SVEPTDYQLG
