PHHY_CUTCT
ID PHHY_CUTCT Reviewed; 665 AA.
AC P15245;
DT 01-APR-1990, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 3.
DT 03-AUG-2022, entry version 128.
DE RecName: Full=Phenol hydroxylase {ECO:0000303|PubMed:4146224};
DE Short=PHHY {ECO:0000303|PubMed:9634698};
DE EC=1.14.13.7 {ECO:0000269|PubMed:11591156, ECO:0000269|PubMed:1429434, ECO:0000269|PubMed:17425111, ECO:0000269|PubMed:2022646, ECO:0000269|PubMed:3203745, ECO:0000269|PubMed:4146224, ECO:0000269|PubMed:7851397, ECO:0000269|PubMed:7858421};
OS Cutaneotrichosporon cutaneum (Yeast) (Trichosporon cutaneum).
OC Eukaryota; Fungi; Dikarya; Basidiomycota; Agaricomycotina; Tremellomycetes;
OC Trichosporonales; Trichosporonaceae; Cutaneotrichosporon.
OX NCBI_TaxID=5554;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PARTIAL PROTEIN SEQUENCE, FUNCTION, AND
RP CATALYTIC ACTIVITY.
RC STRAIN=ATCC 46490;
RX PubMed=1429434; DOI=10.1128/jb.174.22.7112-7120.1992;
RA Kalin M., Neujahr H.Y., Weissmahr R.N., Sejlitz T., Johl R., Fiechter A.,
RA Reiser J.;
RT "Phenol hydroxylase from Trichosporon cutaneum: gene cloning, sequence
RT analysis, and functional expression in Escherichia coli.";
RL J. Bacteriol. 174:7112-7120(1992).
RN [2]
RP PROTEIN SEQUENCE OF 30-64; 299-311 AND 352-377.
RC STRAIN=ATCC 46490;
RX PubMed=2298204; DOI=10.1111/j.1432-1033.1990.tb15298.x;
RA Sejlitz T., Wernstedt C., Engstroem A., Neujahr H.N.;
RT "Amino acid sequences around the pyridoxal-5'-phosphate-binding sites of
RT phenol hydroxylase.";
RL Eur. J. Biochem. 187:225-228(1990).
RN [3]
RP PROTEIN SEQUENCE OF 2-80 AND 662-665.
RX PubMed=1924266;
RA Sejlitz T., Wernstedt C., Hellman U., Neujahr H.Y.;
RT "The N-terminal amino acid sequence of phenol hydroxylase contains a
RT dinucleotide-binding sequence motif.";
RL Protein Seq. Data Anal. 4:21-23(1991).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, AND
RP ACTIVITY REGULATION.
RX PubMed=4146224; DOI=10.1111/j.1432-1033.1973.tb02851.x;
RA Neujahr H.Y., Gaal A.;
RT "Phenol hydroxylase from yeast. Purification and properties of the enzyme
RT from Trichosporon cutaneum.";
RL Eur. J. Biochem. 35:386-400(1973).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=3203745; DOI=10.1016/0014-5793(88)80988-8;
RA Moertberg M., Neujahr H.Y.;
RT "Activation enthalpies and pH dependence of phenol hydroxylase from
RT Trichosporon cutaneum, in vitro and in situ.";
RL FEBS Lett. 242:75-78(1988).
RN [6]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=2022646; DOI=10.1016/s0021-9258(18)92975-3;
RA Taylor M.G., Massey V.;
RT "Kinetic and isotopic studies of the oxidative half-reaction of phenol
RT hydroxylase.";
RL J. Biol. Chem. 266:8291-8301(1991).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=7858421; DOI=10.1006/prep.1994.1073;
RA Waters S., Neujahr H.Y.;
RT "A fermentor culture for production of recombinant phenol hydroxylase.";
RL Protein Expr. Purif. 5:534-540(1994).
RN [8]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=7851397; DOI=10.1111/j.1432-1033.1995.tb20386.x;
RA Peelen S., Rietjens I.M., Boersma M.G., Vervoort J.;
RT "Conversion of phenol derivatives to hydroxylated products by phenol
RT hydroxylase from Trichosporon cutaneum. A comparison of regioselectivity
RT and rate of conversion with calculated molecular orbital substrate
RT characteristics.";
RL Eur. J. Biochem. 227:284-291(1995).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF ASP-55; ARG-282 AND
RP TYR-290.
RX PubMed=11591156; DOI=10.1021/bi010962y;
RA Xu D., Ballou D.P., Massey V.;
RT "Studies of the mechanism of phenol hydroxylase: mutants Tyr289Phe,
RT Asp54Asn, and Arg281Met.";
RL Biochemistry 40:12369-12378(2001).
RN [10]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=R57;
RX PubMed=17425111; DOI=10.1515/znc-2007-1-215;
RA Gerginova M., Manasiev J., Shivarova N., Alexieva Z.;
RT "Influence of various phenolic compounds on phenol hydroxylase activity of
RT a Trichosporon cutaneum strain.";
RL Z. Naturforsch. C 62:83-86(2007).
RN [11]
RP X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS) OF 2-665 IN COMPLEX WITH FAD AND
RP PHENOL, SUBUNIT, AND COFACTOR.
RC STRAIN=ATCC 46490;
RX PubMed=9634698; DOI=10.1016/s0969-2126(98)00062-8;
RA Enroth C., Neujahr H.Y., Schneider G., Lindqvist Y.;
RT "The crystal structure of phenol hydroxylase in complex with FAD and phenol
RT provides evidence for a concerted conformational change in the enzyme and
RT its cofactor during catalysis.";
RL Structure 6:605-617(1998).
RN [12]
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) IN COMPLEX WITH FAD AND PHENOL, AND
RP COFACTOR.
RX PubMed=12925790; DOI=10.1107/s0907444903014902;
RA Enroth C.;
RT "High-resolution structure of phenol hydroxylase and correction of sequence
RT errors.";
RL Acta Crystallogr. D 59:1597-1602(2003).
CC -!- FUNCTION: Hydroxylates phenol to catechol (PubMed:1429434,
CC PubMed:4146224, PubMed:3203745, PubMed:2022646, PubMed:7858421,
CC PubMed:7851397, PubMed:11591156). Phenol is the best substrate, but the
CC enzyme also accepts isomeric diphenols, hydroxyl-, amino-, halogen- or
CC methyl-substituted phenols and, to a lesser degree, cresols
CC (PubMed:4146224, PubMed:17425111, PubMed:7851397).
CC {ECO:0000269|PubMed:11591156, ECO:0000269|PubMed:1429434,
CC ECO:0000269|PubMed:17425111, ECO:0000269|PubMed:2022646,
CC ECO:0000269|PubMed:3203745, ECO:0000269|PubMed:4146224,
CC ECO:0000269|PubMed:7851397, ECO:0000269|PubMed:7858421}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H(+) + NADPH + O2 + phenol = catechol + H2O + NADP(+);
CC Xref=Rhea:RHEA:17061, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:15882, ChEBI:CHEBI:18135,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; EC=1.14.13.7;
CC Evidence={ECO:0000269|PubMed:11591156, ECO:0000269|PubMed:1429434,
CC ECO:0000269|PubMed:17425111, ECO:0000269|PubMed:2022646,
CC ECO:0000269|PubMed:3203745, ECO:0000269|PubMed:4146224,
CC ECO:0000269|PubMed:7851397, ECO:0000269|PubMed:7858421};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:12925790, ECO:0000269|PubMed:4146224,
CC ECO:0000269|PubMed:9634698};
CC -!- ACTIVITY REGULATION: Inhibited by excess phenol (PubMed:4146224). Heavy
CC metals such AsCuSO(4), AgNO(3), or HgCl(2) severely inhibit activity
CC (PubMed:4146224). {ECO:0000269|PubMed:4146224}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=18 uM for phenol {ECO:0000269|PubMed:4146224};
CC KM=71 uM for NADPH {ECO:0000269|PubMed:4146224};
CC KM=53 uM for O(2) {ECO:0000269|PubMed:4146224};
CC pH dependence:
CC Optimum pH is 7.2-7.6. {ECO:0000269|PubMed:3203745,
CC ECO:0000269|PubMed:4146224};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius.
CC {ECO:0000269|PubMed:3203745};
CC -!- PATHWAY: Aromatic compound metabolism; phenol degradation.
CC {ECO:0000269|PubMed:1429434}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:9634698}.
CC -!- SIMILARITY: Belongs to the PheA/TfdB FAD monooxygenase family.
CC {ECO:0000305}.
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DR EMBL; L04488; AAA34202.1; -; mRNA.
DR PIR; S07772; S07772.
DR PDB; 1FOH; X-ray; 2.40 A; A/B/C/D=2-665.
DR PDB; 1PN0; X-ray; 1.70 A; A/B/C/D=1-665.
DR PDBsum; 1FOH; -.
DR PDBsum; 1PN0; -.
DR AlphaFoldDB; P15245; -.
DR SMR; P15245; -.
DR KEGG; ag:AAA34202; -.
DR BioCyc; MetaCyc:MON-14669; -.
DR UniPathway; UPA00728; -.
DR EvolutionaryTrace; P15245; -.
DR GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR GO; GO:0018662; F:phenol 2-monooxygenase activity; IEA:UniProtKB-EC.
DR GO; GO:0019336; P:phenol-containing compound catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.40.30.20; -; 1.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR002938; FAD-bd.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR012941; Phe_hydrox_C_dim_dom.
DR InterPro; IPR038220; PHOX_C_sf.
DR InterPro; IPR036249; Thioredoxin-like_sf.
DR Pfam; PF01494; FAD_binding_3; 1.
DR Pfam; PF07976; Phe_hydrox_dim; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
DR SUPFAM; SSF52833; SSF52833; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Aromatic hydrocarbons catabolism; Direct protein sequencing;
KW FAD; Flavoprotein; Monooxygenase; NADP; Oxidoreductase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:1924266"
FT CHAIN 2..665
FT /note="Phenol hydroxylase"
FT /id="PRO_0000214045"
FT BINDING 18..19
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:12925790,
FT ECO:0000269|PubMed:9634698, ECO:0007744|PDB:1FOH,
FT ECO:0007744|PDB:1PN0"
FT BINDING 43..45
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:12925790,
FT ECO:0000269|PubMed:9634698, ECO:0007744|PDB:1FOH,
FT ECO:0007744|PDB:1PN0"
FT BINDING 51..56
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:12925790,
FT ECO:0000269|PubMed:9634698, ECO:0007744|PDB:1FOH,
FT ECO:0007744|PDB:1PN0"
FT BINDING 55
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:12925790,
FT ECO:0000269|PubMed:9634698, ECO:0007744|PDB:1FOH,
FT ECO:0007744|PDB:1PN0"
FT BINDING 118
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:12925790,
FT ECO:0000269|PubMed:9634698, ECO:0007744|PDB:1FOH,
FT ECO:0007744|PDB:1PN0"
FT BINDING 290
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:12925790,
FT ECO:0000269|PubMed:9634698, ECO:0007744|PDB:1FOH,
FT ECO:0007744|PDB:1PN0"
FT BINDING 290
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:12925790,
FT ECO:0000269|PubMed:9634698, ECO:0007744|PDB:1FOH,
FT ECO:0007744|PDB:1PN0"
FT BINDING 358
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0007744|PDB:1FOH, ECO:0007744|PDB:1PN0"
FT BINDING 368..372
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:12925790,
FT ECO:0000269|PubMed:9634698, ECO:0007744|PDB:1FOH,
FT ECO:0007744|PDB:1PN0"
FT MUTAGEN 55
FT /note="D->N: Leads to a significant slower initial step of
FT the oxidative half-reaction."
FT /evidence="ECO:0000269|PubMed:11591156"
FT MUTAGEN 282
FT /note="R->M: Leads to a significant slower initial step of
FT the oxidative half-reaction."
FT /evidence="ECO:0000269|PubMed:11591156"
FT MUTAGEN 290
FT /note="Y->F: Leads to a slightly higher redox potential but
FT is reduced by NADPH much slower."
FT /evidence="ECO:0000269|PubMed:11591156"
FT STRAND 4..14
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 18..33
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 39..42
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 44..47
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 59..66
FT /evidence="ECO:0007829|PDB:1PN0"
FT TURN 67..69
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 71..75
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 83..89
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 95..104
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 118..133
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 142..150
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 152..154
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 163..169
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 172..174
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 186..188
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 192..200
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 214..224
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 231..236
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 241..257
FT /evidence="ECO:0007829|PDB:1PN0"
FT TURN 261..264
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 265..270
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 272..274
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 276..281
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 287..293
FT /evidence="ECO:0007829|PDB:1PN0"
FT TURN 305..307
FT /evidence="ECO:0007829|PDB:1FOH"
FT HELIX 310..321
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 327..343
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 347..349
FT /evidence="ECO:0007829|PDB:1PN0"
FT TURN 350..352
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 353..355
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 357..359
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 370..389
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 395..399
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 400..424
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 429..432
FT /evidence="ECO:0007829|PDB:1FOH"
FT STRAND 435..437
FT /evidence="ECO:0007829|PDB:1FOH"
FT HELIX 439..453
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 473..475
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 489..492
FT /evidence="ECO:0007829|PDB:1PN0"
FT TURN 493..496
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 497..500
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 501..504
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 511..518
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 523..537
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 542..546
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 555..565
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 572..574
FT /evidence="ECO:0007829|PDB:1PN0"
FT TURN 577..580
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 587..591
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 596..598
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 603..607
FT /evidence="ECO:0007829|PDB:1PN0"
FT TURN 611..613
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 615..619
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 623..629
FT /evidence="ECO:0007829|PDB:1PN0"
FT HELIX 634..642
FT /evidence="ECO:0007829|PDB:1PN0"
FT STRAND 649..651
FT /evidence="ECO:0007829|PDB:1PN0"
SQ SEQUENCE 665 AA; 75162 MW; 1401BE35D38BFB71 CRC64;
MTKYSESYCD VLIVGAGPAG LMAARVLSEY VRQKPDLKVR IIDKRSTKVY NGQADGLQCR
TLESLKNLGL ADKILSEAND MSTIALYNPD ENGHIRRTDR IPDTLPGISR YHQVVLHQGR
IERHILDSIA EISDTRIKVE RPLIPEKMEI DSSKAEDPEA YPVTMTLRYM SDHESTPLQF
GHKTENSLFH SNLQTQEEED ANYRLPEGKE AGEIETVHCK YVIGCDGGHS WVRRTLGFEM
IGEQTDYIWG VLDAVPASNF PDIRSPCAIH SAESGSIMII PRENNLVRFY VQLQARAEKG
GRVDRTKFTP EVVIANAKKI FHPYTFDVQQ LDWFTAYHIG QRVTEKFSKD ERVFIAGDAC
HTHSPKAGQG MNTSMMDTYN LGWKLGLVLT GRAKRDILKT YEEERHAFAQ ALIDFDHQFS
RLFSGRPAKD VADEMGVSMD VFKEAFVKGN EFASGTAINY DENLVTDKKS SKQELAKNCV
VGTRFKSQPV VRHSEGLWMH FGDRLVTDGR FRIIVFAGKA TDATQMSRIK KFSAYLDSEN
SVISLYTPKV SDRNSRIDVI TIHSCHRDDI EMHDFPAPAL HPKWQYDFIY ADCDSWHHPH
PKSYQAWGVD ETKGAVVVVR PDGYTSLVTD LEGTAEIDRY FSGILVEPKE KSGAQTEADW
TKSTA
