P2OX_PHLGI
ID P2OX_PHLGI Reviewed; 622 AA.
AC Q6UG02;
DT 07-JUN-2005, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 69.
DE RecName: Full=Pyranose 2-oxidase;
DE Short=P2Ox;
DE Short=POD;
DE Short=POx;
DE Short=PROD;
DE Short=Pyranose oxidase;
DE EC=1.1.3.10;
DE AltName: Full=FAD-oxidoreductase;
DE AltName: Full=Glucose 2-oxidase;
DE AltName: Full=Pyranose:oxygen 2-oxidoreductase;
DE Flags: Precursor;
GN Name=p2ox; Synonyms=poxB;
OS Phlebiopsis gigantea (White-rot fungus) (Peniophora gigantea).
OC Eukaryota; Fungi; Dikarya; Basidiomycota; Agaricomycotina; Agaricomycetes;
OC Polyporales; Phanerochaetaceae; Phlebiopsis.
OX NCBI_TaxID=82310;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 29-52, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND MUTAGENESIS OF HIS-167; LYS-312 AND GLU-540.
RC STRAIN=DSM 13218;
RX PubMed=15660220; DOI=10.1007/s00253-004-1813-1;
RA Bastian S., Rekowski M.J., Witte K., Heckmann-Pohl D.M., Giffhorn F.;
RT "Engineering of pyranose 2-oxidase from Peniophora gigantea towards
RT improved thermostability and catalytic efficiency.";
RL Appl. Microbiol. Biotechnol. 67:654-663(2005).
RN [2]
RP BIOPHYSICOCHEMICAL PROPERTIES, TETRAMERIZATION, AND FAD-BINDING.
RX PubMed=8319689; DOI=10.1111/j.1432-1033.1993.tb17982.x;
RA Danneel H.-J., Roessner E., Zeeck A., Giffhorn F.;
RT "Purification and characterization of a pyranose oxidase from the
RT basidiomycete Peniophora gigantea and chemical analyses of its reaction
RT products.";
RL Eur. J. Biochem. 214:795-802(1993).
CC -!- FUNCTION: Catalyzes the oxidation of various aldopyranoses and
CC disaccharides on carbon-2 to the corresponding 2-keto sugars
CC concomitant with the reduction of O(2) to H(2)O(2). Plays an important
CC role in lignin degradation of wood rot fungi by supplying the essential
CC cosubstrate H(2)O(2) for the ligninolytic peroxidases, lignin
CC peroxidase and manganese-dependent peroxidase. The preferred substrate
CC is D-glucose which is converted to 2-dehydro-D-glucose, an intermediate
CC of a secondary metabolic pathway leading to the antibiotic
CC cortalcerone. Acts also on D-xylose, together with D-glucose the major
CC sugars derived from wood, on L-sorbose, D-galactose and 1,5-
CC anhydroglucitol, a diagnostic marker of diabetes mellitus.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=D-glucose + O2 = 2-dehydro-D-glucose + H2O2;
CC Xref=Rhea:RHEA:10552, ChEBI:CHEBI:4167, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16240, ChEBI:CHEBI:16609; EC=1.1.3.10;
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:8319689};
CC Note=Binds 1 FAD covalently per subunit. {ECO:0000269|PubMed:8319689};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.65 mM for O(2) {ECO:0000269|PubMed:15660220,
CC ECO:0000269|PubMed:8319689};
CC KM=1.1 mM for D-glucose {ECO:0000269|PubMed:15660220,
CC ECO:0000269|PubMed:8319689};
CC KM=11.3 mM for 2-deoxy-D-glucose {ECO:0000269|PubMed:15660220,
CC ECO:0000269|PubMed:8319689};
CC KM=289 mM for methyl-beta-D-glucoside {ECO:0000269|PubMed:15660220,
CC ECO:0000269|PubMed:8319689};
CC KM=29.4 mM for D-xylose {ECO:0000269|PubMed:15660220,
CC ECO:0000269|PubMed:8319689};
CC KM=50 mM for L-sorbose {ECO:0000269|PubMed:15660220,
CC ECO:0000269|PubMed:8319689};
CC KM=8.2 mM for D-galactose {ECO:0000269|PubMed:15660220,
CC ECO:0000269|PubMed:8319689};
CC KM=55.7 mM for D-allose {ECO:0000269|PubMed:15660220,
CC ECO:0000269|PubMed:8319689};
CC Vmax=10.4 umol/min/mg enzyme with D-glucose as substrate
CC {ECO:0000269|PubMed:15660220, ECO:0000269|PubMed:8319689};
CC Vmax=4.5 umol/min/mg enzyme with 2-deoxy-D-glucose as substrate
CC {ECO:0000269|PubMed:15660220, ECO:0000269|PubMed:8319689};
CC Vmax=2.9 umol/min/mg enzyme with methyl-beta-D-glucoside as substrate
CC {ECO:0000269|PubMed:15660220, ECO:0000269|PubMed:8319689};
CC Vmax=4.3 umol/min/mg enzyme with D-xylose as substrate
CC {ECO:0000269|PubMed:15660220, ECO:0000269|PubMed:8319689};
CC Vmax=7.9 umol/min/mg enzyme with L-sorbose as substrate
CC {ECO:0000269|PubMed:15660220, ECO:0000269|PubMed:8319689};
CC Vmax=0.5 umol/min/mg enzyme with D-galactose as substrate
CC {ECO:0000269|PubMed:15660220, ECO:0000269|PubMed:8319689};
CC Vmax=3.7 umol/min/mg enzyme with D-allose as substrate
CC {ECO:0000269|PubMed:15660220, ECO:0000269|PubMed:8319689};
CC pH dependence:
CC Optimum pH is 4.5-6.0. Active from pH 4 to 7.5. Stable from pH 4 to
CC 11. {ECO:0000269|PubMed:15660220, ECO:0000269|PubMed:8319689};
CC Temperature dependence:
CC Optimum temperature is 44 degrees Celsius. Active from 30 to 55
CC degrees Celsius. Thermostable for 30 minutes up to 50 degrees
CC Celsius. {ECO:0000269|PubMed:15660220, ECO:0000269|PubMed:8319689};
CC -!- SUBUNIT: Homotetramer.
CC -!- SUBCELLULAR LOCATION: Periplasm {ECO:0000250}. Note=Hyphal periplasmic
CC space. {ECO:0000250}.
CC -!- PTM: Not glycosylated.
CC -!- SIMILARITY: Belongs to the GMC oxidoreductase family. {ECO:0000305}.
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DR EMBL; AY370876; AAQ72486.1; -; mRNA.
DR AlphaFoldDB; Q6UG02; -.
DR SMR; Q6UG02; -.
DR CAZy; AA3; Auxiliary Activities 3.
DR BRENDA; 1.1.3.10; 4657.
DR SABIO-RK; Q6UG02; -.
DR GO; GO:0042597; C:periplasmic space; IEA:UniProtKB-SubCell.
DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IEA:InterPro.
DR GO; GO:0050233; F:pyranose oxidase activity; IEA:UniProtKB-EC.
DR Gene3D; 3.50.50.60; -; 2.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR000172; GMC_OxRdtase_N.
DR InterPro; IPR007867; GMC_OxRtase_C.
DR InterPro; IPR012814; P2OX.
DR Pfam; PF05199; GMC_oxred_C; 1.
DR Pfam; PF00732; GMC_oxred_N; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
DR TIGRFAMs; TIGR02462; pyranose_ox; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; FAD; Flavoprotein; Oxidoreductase; Periplasm;
KW Signal.
FT SIGNAL 1..28
FT /evidence="ECO:0000250"
FT PROPEP 29..37
FT /evidence="ECO:0000250"
FT /id="PRO_0000012350"
FT CHAIN 38..622
FT /note="Pyranose 2-oxidase"
FT /id="PRO_0000012351"
FT ACT_SITE 546
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:E4QP00"
FT ACT_SITE 591
FT /evidence="ECO:0000250"
FT BINDING 449
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 451
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT MOD_RES 167
FT /note="Tele-8alpha-FAD histidine"
FT /evidence="ECO:0000305"
FT MUTAGEN 167
FT /note="H->C: Decreases activity by 90%."
FT /evidence="ECO:0000269|PubMed:15660220"
FT MUTAGEN 312
FT /note="K->E: In P2OxB2H; increases the catalytic efficiency
FT 5.3-fold for D-glucose, 2-fold for methyl-beta-D-glucoside,
FT 59.9-fold for D-xylose, 69-fold for L-sorbose and 4.8-fold
FT for D-galactose; when associated with K-540."
FT /evidence="ECO:0000269|PubMed:15660220"
FT MUTAGEN 540
FT /note="E->K: In P2OxB1; increases thermostability up to 70
FT degrees Celsius and enhances pH stability in alkaline
FT solution. Increases the catalytic efficiency 3.1-fold for
FT D-xylose and 7.3-fold for L-sorbose, mainly by lowering the
FT Km values for these two substrates to 6.6 mM and 5.4 mM,
FT respectively. In P2OxB2H; increases the catalytic
FT efficiency 5.3-fold for D-glucose, 2-fold for methyl-beta-
FT D-glucoside, 59.9-fold for D-xylose, 69-fold for L-sorbose
FT and 4.8-fold for D-galactose; when associated with K-312."
FT /evidence="ECO:0000269|PubMed:15660220"
SQ SEQUENCE 622 AA; 68902 MW; 3F69FC03DC6BF134 CRC64;
MSASSSDPFH SFAKTSFTSK AAKRATAHSL PPLPGPGDLP PGMNVEYDVA IVGSGPIGST
YARELVEAGF NVAMFEIGEI DSGLKIGSHK KNTVEYQKNI DKFVNVIQGQ LMPVSVPVNT
MVVDTLSPAS WQASTFFVRN GANPEQDPLR NLSGQAVTRV VGGMSTHWTC ATPRFEKLQR
PLLVKNDPVA DDAEWDRLYK KAESYFKTGT TQFAESIRHN LVLKKLQEEY KGVRDFQQIP
LAATRQSPTF VEWSSAHTVF DLENRPNKDA PKQRFNLFPA VACTSVRRND ANSEIIGLDV
RDLHGGKSIT IKAKVYILTA GAVHNAQLLA ASGFGQLGRP DPAKPLPSLL PYLGTHITEQ
TLVFCQTVMS TELINSVTAD MTIVGKPGDP DYSVTYTSGS PNNKHPDWWN EKVKKHMMDH
QEDPLPIPFE DPEPQVTTLF KASHPWHTQI HRDAFSYGAV QQSIDSRLIV DWRFFGRTEP
KEENKLWFSD KITDAYNLPQ PTFDFRFPGG REAEDMMTDM CVMSAKIGGF LPGSYPQFME
PGLVLHLGGT HRMGFDEKAD KCCVDTDSRV FGFKNLFLGG CGNIPTAYAA NPTLTAMSLA
IKSCEYIKKN FEPSPNPVKH HN
