HMFO_METS6
ID HMFO_METS6 Reviewed; 531 AA.
AC E4QP00;
DT 01-APR-2015, integrated into UniProtKB/Swiss-Prot.
DT 08-FEB-2011, sequence version 1.
DT 03-AUG-2022, entry version 59.
DE RecName: Full=5-(hydroxymethyl)furfural oxidase {ECO:0000305};
DE EC=1.1.3.47 {ECO:0000269|PubMed:24271187, ECO:0000269|PubMed:24802551};
DE AltName: Full=5-hydroxymethylfurfural oxidase {ECO:0000303|PubMed:24271187};
DE Short=HMFO {ECO:0000303|PubMed:24271187};
DE AltName: Full=Thiol oxidase {ECO:0000305|PubMed:25284255};
DE EC=1.8.3.- {ECO:0000269|PubMed:25284255};
GN OrderedLocusNames=MPQ_0130 {ECO:0000312|EMBL:ADQ83320.1};
OS Methylovorus sp. (strain MP688).
OC Bacteria; Proteobacteria; Betaproteobacteria; Nitrosomonadales;
OC Methylophilaceae; Methylovorus; unclassified Methylovorus.
OX NCBI_TaxID=887061;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=MP688;
RX PubMed=21148725; DOI=10.1128/jb.01431-10;
RA Xiong X.H., Zhi J.J., Yang L., Wang J.H., Zhao Y., Wang X., Cui Y.J.,
RA Dong F., Li M.X., Yang Y.X., Wei N., An J.J., Du B.H., Liang L.,
RA Zhang J.S., Zhou W., Cheng S.F., He T., Wang L., Chen H.P., Liu D.S.,
RA Zhang W.C.;
RT "Complete genome sequence of the bacterium Methylovorus sp. strain MP688, a
RT high-level producer of pyrroloquinolone quinone.";
RL J. Bacteriol. 193:1012-1013(2011).
RN [2]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=24802551; DOI=10.1002/anie.201402904;
RA Dijkman W.P., Groothuis D.E., Fraaije M.W.;
RT "Enzyme-catalyzed oxidation of 5-hydroxymethylfurfural to furan-2,5-
RT dicarboxylic acid.";
RL Angew. Chem. Int. Ed. Engl. 53:6515-6518(2014).
RN [3]
RP FUNCTION AS A THIOL OXIDASE, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RC STRAIN=MP688;
RX PubMed=25284255; DOI=10.1002/anie.201407520;
RA Ewing T.A., Dijkman W.P., Vervoort J.M., Fraaije M.W., van Berkel W.J.;
RT "The oxidation of thiols by flavoprotein oxidases: a biocatalytic route to
RT reactive thiocarbonyls.";
RL Angew. Chem. Int. Ed. Engl. 53:13206-13209(2014).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP ACTIVE SITE, AND MUTAGENESIS OF HIS-467.
RC STRAIN=MP688;
RX PubMed=24271187; DOI=10.1128/aem.03740-13;
RA Dijkman W.P., Fraaije M.W.;
RT "Discovery and characterization of a 5-hydroxymethylfurfural oxidase from
RT Methylovorus sp. strain MP688.";
RL Appl. Environ. Microbiol. 80:1082-1090(2014).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF WILD-TYPE AND MUTANT ALA-467 IN
RP COMPLEXES WITH FAD, COFACTOR, BIOTECHNOLOGY, MUTAGENESIS OF VAL-101;
RP MET-103; VAL-367; TRP-369; VAL-465; TRP-466 AND ASN-511, ACTIVE SITE, AND
RP SUBUNIT.
RC STRAIN=MP688;
RX DOI=10.1021/ACSCATAL.5B00031;
RA Dijkman W.P., Binda C., Fraaije M.W., Mattevi A.;
RT "Structure-based enzyme tailoring of 5-hydroxymethylfurfural oxidase.";
RL ACS Catal. 5:1833-1839(2015).
CC -!- FUNCTION: Involved in the degradation and detoxification of 5-
CC (hydroxymethyl)furfural (HMF) by mediating its oxidation to furan-2,5-
CC dicarboxylate (FDCA), a biobased platform chemical for the production
CC of polymers. Active with a wide range of aromatic and aliphatic primary
CC alcohols and aldehydes: acts on alcohol groups and requires the
CC spontaneous hydration of aldehyde groups for their oxidation
CC (PubMed:24271187, PubMed:24802551). To a lesser extent, is also able to
CC catalyze the oxidation of thiols that are structurally similar to its
CC alcohol substrates, yielding the corresponding thiocarbonyls
CC (PubMed:25284255). {ECO:0000269|PubMed:24271187,
CC ECO:0000269|PubMed:24802551, ECO:0000269|PubMed:25284255}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=5-hydroxymethylfurfural + 2 H2O + 3 O2 = 2,5-dicarboxyfuran +
CC 2 H(+) + 3 H2O2; Xref=Rhea:RHEA:32683, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:83389, ChEBI:CHEBI:412516; EC=1.1.3.47;
CC Evidence={ECO:0000269|PubMed:24271187, ECO:0000269|PubMed:24802551};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=benzylthiol + O2 = benzothialdehyde + H2O2;
CC Xref=Rhea:RHEA:53792, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:137674, ChEBI:CHEBI:137675;
CC Evidence={ECO:0000269|PubMed:25284255};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:24271187, ECO:0000305|Ref.5};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.4 mM for 5-(hydroxymethyl)furfural
CC {ECO:0000269|PubMed:24271187};
CC KM=1.5 mM for 1,4-benzenedimethanol {ECO:0000269|PubMed:24271187};
CC KM=1.4 mM for 1,3-benzenedimethanol {ECO:0000269|PubMed:24271187};
CC KM=1.3 mM for benzyl alcohol {ECO:0000269|PubMed:24271187};
CC KM=0.3 mM for 4-hydroxybenzyl alcohol {ECO:0000269|PubMed:24271187};
CC KM=1.44 mM for 4-aminobenzyl alcohol {ECO:0000269|PubMed:24271187};
CC KM=0.08 mM for 4-chlorobenzyl alcohol {ECO:0000269|PubMed:24271187};
CC KM=0.07 mM for cinnamyl alcohol {ECO:0000269|PubMed:24271187};
CC KM=0.59 mM for 2,4-hexadien-1-ol {ECO:0000269|PubMed:24271187};
CC KM=0.73 mM for vanillyl alcohol {ECO:0000269|PubMed:24271187};
CC KM=3.5 mM for phenylmethanethiol {ECO:0000269|PubMed:25284255};
CC KM=15 mM for (4-nitrophenyl)methanethiol
CC {ECO:0000269|PubMed:25284255};
CC KM=0.078 mM for (4-nitrophenyl)methanol
CC {ECO:0000269|PubMed:25284255};
CC Note=kcat is 9.9 sec(-1) with 5-(hydroxymethyl)furfural as substrate.
CC kcat is 21.1 sec(-1) with 1,4-benzenedimethanol as substrate. kcat is
CC 14.0 sec(-1) with 1,3-benzenedimethanol as substrate. kcat is 13.5
CC sec(-1) with benzyl alcohol as substrate. kcat is 7.2 sec(-1) with 4-
CC hydroxybenzyl alcohol as substrate. kcat is 17.1 sec(-1) with 4-
CC aminobenzyl alcohol as substrate. kcat is 9.5 sec(-1) with 4-
CC chlorobenzyl alcohol as substrate. kcat is 17.0 sec(-1) with cinnamyl
CC alcohol as substrate. kcat is 13.3 sec(-1) with 2,4-hexadien-1-ol as
CC substrate. kcat is 21.0 sec(-1) with vanillyl alcohol as substrate
CC (PubMed:24271187). kcat is 2.1 sec(-1) with phenylmethanethiol as
CC substrate. kcat is 3.0 sec(-1) with (4-nitrophenyl)phenylmethanethiol
CC as substrate. kcat is 4.5 sec(-1) with (4-nitrophenyl)methanol as
CC substrate (PubMed:25284255). {ECO:0000269|PubMed:24271187,
CC ECO:0000269|PubMed:25284255};
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:24271187};
CC Temperature dependence:
CC Optimum temperature is 55 degrees Celsius.
CC {ECO:0000269|PubMed:24271187};
CC -!- SUBUNIT: Monomer. {ECO:0000269|Ref.5}.
CC -!- BIOTECHNOLOGY: Because 5-hydroxymethylfurfural can be derived from
CC sugars, the enzyme is of particular interest for the production of
CC biobased plastic materials in an environmentally friendly industrial
CC process. {ECO:0000305|Ref.5}.
CC -!- MISCELLANEOUS: At pH 7.0, phenylmethanethiol is oxidized to the
CC corresponding aromatic thioaldehyde, benzothialdehyde, and no formation
CC of 1,2-dibenzyldisulfane is observed, demonstrating that HMFO does not
CC catalyze the formation of disulfide bonds. At pH 8.0, two products are
CC formed, benzothialdehyde and benzaldehyde; this suggests that the
CC thioaldehyde is slowly hydrated, yielding the aldehyde as the final
CC product. {ECO:0000269|PubMed:25284255}.
CC -!- SIMILARITY: Belongs to the GMC oxidoreductase family. {ECO:0000305}.
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DR EMBL; CP002252; ADQ83320.1; -; Genomic_DNA.
DR RefSeq; WP_013440946.1; NC_014733.1.
DR PDB; 4UDP; X-ray; 1.90 A; A/B=1-531.
DR PDB; 4UDQ; X-ray; 1.60 A; A/B=1-531.
DR PDB; 4UDR; X-ray; 1.60 A; A/B=1-531.
DR PDB; 6F97; X-ray; 1.90 A; A/B=1-531.
DR PDBsum; 4UDP; -.
DR PDBsum; 4UDQ; -.
DR PDBsum; 4UDR; -.
DR PDBsum; 6F97; -.
DR AlphaFoldDB; E4QP00; -.
DR SMR; E4QP00; -.
DR KEGG; mep:MPQ_0130; -.
DR HOGENOM; CLU_002865_7_1_4; -.
DR OMA; WHYPVPE; -.
DR BioCyc; MetaCyc:MON-17167; -.
DR BRENDA; 1.1.3.47; 13773.
DR GO; GO:0071949; F:FAD binding; IDA:UniProtKB.
DR GO; GO:0016670; F:oxidoreductase activity, acting on a sulfur group of donors, oxygen as acceptor; IDA:UniProtKB.
DR GO; GO:0016899; F:oxidoreductase activity, acting on the CH-OH group of donors, oxygen as acceptor; IDA:UniProtKB.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR012132; GMC_OxRdtase.
DR InterPro; IPR000172; GMC_OxRdtase_N.
DR InterPro; IPR007867; GMC_OxRtase_C.
DR PANTHER; PTHR11552; PTHR11552; 2.
DR Pfam; PF05199; GMC_oxred_C; 1.
DR Pfam; PF00732; GMC_oxred_N; 1.
DR PIRSF; PIRSF000137; Alcohol_oxidase; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; FAD; Flavoprotein; Oxidoreductase.
FT CHAIN 1..531
FT /note="5-(hydroxymethyl)furfural oxidase"
FT /id="PRO_0000432710"
FT ACT_SITE 467
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:24271187, ECO:0000305|Ref.5"
FT BINDING 15..16
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000305|Ref.5, ECO:0007744|PDB:4UDP,
FT ECO:0007744|PDB:4UDQ, ECO:0007744|PDB:4UDR"
FT BINDING 36..37
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000305|Ref.5, ECO:0007744|PDB:4UDP,
FT ECO:0007744|PDB:4UDQ, ECO:0007744|PDB:4UDR"
FT BINDING 68
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000305|Ref.5, ECO:0007744|PDB:4UDP,
FT ECO:0007744|PDB:4UDQ, ECO:0007744|PDB:4UDR"
FT BINDING 94
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000305|Ref.5, ECO:0007744|PDB:4UDP,
FT ECO:0007744|PDB:4UDQ, ECO:0007744|PDB:4UDR"
FT BINDING 98
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000305|Ref.5, ECO:0007744|PDB:4UDP,
FT ECO:0007744|PDB:4UDQ, ECO:0007744|PDB:4UDR"
FT BINDING 102..105
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000305|Ref.5, ECO:0007744|PDB:4UDP,
FT ECO:0007744|PDB:4UDQ, ECO:0007744|PDB:4UDR"
FT BINDING 233
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000305|Ref.5, ECO:0007744|PDB:4UDP,
FT ECO:0007744|PDB:4UDQ, ECO:0007744|PDB:4UDR"
FT BINDING 466
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0007744|PDB:4UDR"
FT BINDING 501
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000305|Ref.5, ECO:0007744|PDB:4UDP,
FT ECO:0007744|PDB:4UDQ, ECO:0007744|PDB:4UDR"
FT BINDING 512..513
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000305|Ref.5, ECO:0007744|PDB:4UDP,
FT ECO:0007744|PDB:4UDQ, ECO:0007744|PDB:4UDR"
FT MUTAGEN 101
FT /note="V->H: Abolishes activity."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 103
FT /note="M->A: 16-fold reduction in catalytic efficiency on
FT vanillyl alcohol."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 367
FT /note="V->K: 1.6-fold reduction in catalytic efficiency on
FT vanillyl alcohol. Shows significantly improved activity on
FT the aldehyde 5-formyl-2-furancarboxylate, which results in
FT a better 5-hydroxymethylfurfural to 2,5-furandicarboxylate
FT conversion."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 367
FT /note="V->R: 1.4-fold reduction in catalytic efficiency on
FT vanillyl alcohol. Shows significantly improved activity on
FT the aldehyde 5-formyl-2-furancarboxylate, which results in
FT a better 5-hydroxymethylfurfural to 2,5-furandicarboxylate
FT conversion. Displays a catalytic efficiency toward 5-
FT formyl-2-furancarboxylate that is over 1000-fold higher
FT than that for wild-type; when associated with F-466."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 369
FT /note="W->A: 7.5-fold reduction in catalytic efficiency on
FT vanillyl alcohol."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 465
FT /note="V->A: 18-fold reduction in catalytic efficiency on
FT vanillyl alcohol."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 466
FT /note="W->A: 39-fold reduction in catalytic efficiency on
FT vanillyl alcohol. In contrast to wild-type, is active on
FT secondary alcohols, such as (S)-1-phenylethanol, and is
FT strictly enantionselective as this mutant has no activity
FT on (R)-1-phenylethanol. Shows increased activity on the
FT aldehyde 5-formyl-2-furancarboxylate."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 466
FT /note="W->F: 3.4-fold reduction in catalytic efficiency on
FT vanillyl alcohol. In contrast to wild-type, is active on
FT secondary alcohols, such as (S)-1-phenylethanol, and is
FT strictly enantionselective as this mutant has no activity
FT on (R)-1-phenylethanol. Shows increased activity on the
FT aldehyde 5-formyl-2-furancarboxylate. Displays a catalytic
FT efficiency toward 5-formyl-2-furancarboxylate that is over
FT 1000-fold higher than that for wild-type; when associated
FT with R-367."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 467
FT /note="H->A: Abolishes activity."
FT /evidence="ECO:0000269|PubMed:24271187"
FT MUTAGEN 511
FT /note="N->A: 53-fold reduction in catalytic efficiency on
FT vanillyl alcohol."
FT /evidence="ECO:0000269|Ref.5"
FT STRAND 6..11
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 15..25
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 32..35
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 37..39
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 43..45
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 48..51
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 52..55
FT /evidence="ECO:0007829|PDB:4UDP"
FT HELIX 58..61
FT /evidence="ECO:0007829|PDB:4UDQ"
FT TURN 63..66
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 72..76
FT /evidence="ECO:0007829|PDB:4UDQ"
FT TURN 95..97
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 98..101
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 111..119
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 127..137
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 138..140
FT /evidence="ECO:0007829|PDB:4UDQ"
FT TURN 141..145
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 151..158
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 162..164
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 167..178
FT /evidence="ECO:0007829|PDB:4UDQ"
FT TURN 187..189
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 192..196
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 210..214
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 217..220
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 225..228
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 230..239
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 242..250
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 253..264
FT /evidence="ECO:0007829|PDB:4UDQ"
FT TURN 268..270
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 271..278
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 283..288
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 294..296
FT /evidence="ECO:0007829|PDB:4UDQ"
FT TURN 298..301
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 303..305
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 308..315
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 318..324
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 336..339
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 351..358
FT /evidence="ECO:0007829|PDB:4UDQ"
FT TURN 359..362
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 363..372
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 377..380
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 382..384
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 391..396
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 400..418
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 420..423
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 430..432
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 434..436
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 444..447
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 451..461
FT /evidence="ECO:0007829|PDB:4UDQ"
FT STRAND 488..490
FT /evidence="ECO:0007829|PDB:6F97"
FT STRAND 493..498
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 501..503
FT /evidence="ECO:0007829|PDB:4UDQ"
FT HELIX 513..528
FT /evidence="ECO:0007829|PDB:4UDQ"
SQ SEQUENCE 531 AA; 57014 MW; 111A92125A89F295 CRC64;
MTDTIFDYVI VGGGTAGSVL ANRLSARPEN RVLLIEAGID TPENNIPPEI HDGLRPWLPR
LSGDKFFWPN LTIHRAAEHP GITREPQFYE QGRLLGGGSS VNMVVSNRGL PRDYDEWQAL
GADGWDWQGV LPYFIKTERD ADYGDDPLHG NAGPIPIGRV DSRHWSDFTV AATQALEAAG
LPNIHDQNAR FDDGYFPPAF TLKGEERFSA ARGYLDASVR VRPNLSLWTE SRVLKLLTTG
NAITGVSVLR GRETLQVQAR EVILTAGALQ SPAILLRTGI GPAADLHALG IPVLADRPGV
GRNLWEHSSI GVVAPLTEQA RADASTGKAG SRHQLGIRAS SGVDPATPSD LFLHIGADPV
SGLASAVFWV NKPSSTGWLK LKDADPFSYP DVDFNLLSDP RDLGRLKAGL RLITHYFAAP
SLAKYGLALA LSRFAAPQPG GPLLNDLLQD EAALERYLRT NVGGVWHASG TARIGRADDS
QAVVDKAGRV YGVTGLRVAD ASIMPTVPTA NTNLPTLMLA EKIADAILTQ A
