ID   MDLB_PSEPU              Reviewed;         393 AA.
AC   P20932;
DT   01-FEB-1991, integrated into UniProtKB/Swiss-Prot.
DT   01-FEB-1991, sequence version 1.
DT   02-SEP-2026, entry version 129.
DE   RecName: Full=(S)-mandelate dehydrogenase {ECO:0000303|PubMed:10493804, ECO:0000303|PubMed:11502180, ECO:0000303|PubMed:2271624};
DE            Short=MDH {ECO:0000303|PubMed:10493804, ECO:0000303|PubMed:2271624};
DE            EC=1.1.99.31;
DE   AltName: Full=L(+)-mandelate dehydrogenase;
GN   Name=mdlB {ECO:0000303|PubMed:2271624};
OS   Pseudomonas putida (Arthrobacter siderocapsulatus).
OC   Bacteria; Pseudomonadati; Pseudomonadota; Gammaproteobacteria;
OC   Pseudomonadales; Pseudomonadaceae; Pseudomonas.
OX   NCBI_TaxID=303;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-30, FUNCTION, AND
RP   PATHWAY.
RC   STRAIN=ATCC 12633 / DSM 291 / JCM 13063 / CCUG 12690 / LMG 2257 / NBRC
RC   14164 / NCIMB 9494 / NCTC 10936 / VKM B-2187 / Stanier 90;
RX   PubMed=2271624; DOI=10.1021/bi00494a015;
RA   Tsou A.Y., Ransom S.C., Gerlt J.A., Buechter D.D., Babbitt P.C.,
RA   Kenyon G.L.;
RT   "Mandelate pathway of Pseudomonas putida: sequence relationships involving
RT   mandelate racemase, (S)-mandelate dehydrogenase, and benzoylformate
RT   decarboxylase and expression of benzoylformate decarboxylase in Escherichia
RT   coli.";
RL   Biochemistry 29:9856-9862(1990).
RN   [2]
RP   FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP   SUBSTRATE SPECIFICITY, AND SUBCELLULAR LOCATION.
RC   STRAIN=ATCC 12633 / DSM 291 / JCM 13063 / CCUG 12690 / LMG 2257 / NBRC
RC   14164 / NCIMB 9494 / NCTC 10936 / VKM B-2187 / Stanier 90;
RX   PubMed=10493804; DOI=10.1021/bi990996u;
RA   Xu Y., Mitra B.;
RT   "A highly active, soluble mutant of the membrane-associated (S)-mandelate
RT   dehydrogenase from Pseudomonas putida.";
RL   Biochemistry 38:12367-12376(1999).
RN   [3] {ECO:0007744|PDB:1HUV}
RP   X-RAY CRYSTALLOGRAPHY (2.15 ANGSTROMS) OF A CHIMERIC MUTANT IN COMPLEX WITH
RP   FMN, COFACTOR, AND SUBUNIT.
RX   PubMed=11502180; DOI=10.1021/bi010938k;
RA   Sukumar N., Xu Y., Gatti D.L., Mitra B., Mathews F.S.;
RT   "Structure of an active soluble mutant of the membrane-associated (S)-
RT   mandelate dehydrogenase.";
RL   Biochemistry 40:9870-9878(2001).
RN   [4] {ECO:0007744|PDB:1P4C, ECO:0007744|PDB:1P5B}
RP   X-RAY CRYSTALLOGRAPHY (1.35 ANGSTROMS) OF A CHIMERIC MUTANT IN COMPLEX WITH
RP   FMN, COFACTOR, AND SUBUNIT.
RX   PubMed=14604988; DOI=10.1074/jbc.m310049200;
RA   Sukumar N., Dewanti A.R., Mitra B., Mathews F.S.;
RT   "High resolution structures of an oxidized and reduced flavoprotein. The
RT   water switch in a soluble form of (S)-mandelate dehydrogenase.";
RL   J. Biol. Chem. 279:3749-3757(2004).
RN   [5] {ECO:0007744|PDB:2A7P, ECO:0007744|PDB:2A85, ECO:0007744|PDB:3GIY}
RP   X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF MUTANT ALA-81 OF A CHIMERIC
RP   MUTANT IN COMPLEXES WITH (2S)-2-HYDROXYOCTANOATE; 3-INDOLELACTATE AND FMN,
RP   FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, AND
RP   MUTAGENESIS OF GLY-81.
RX   PubMed=19465768; DOI=10.1107/s0907444909010270;
RA   Sukumar N., Dewanti A., Merli A., Rossi G.L., Mitra B., Mathews F.S.;
RT   "Structures of the G81A mutant form of the active chimera of (S)-mandelate
RT   dehydrogenase and its complex with two of its substrates.";
RL   Acta Crystallogr. D 65:543-552(2009).
RN   [6] {ECO:0007744|PDB:6BFG}
RP   X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH FMN, AND COFACTOR.
RX   PubMed=30071260; DOI=10.1016/j.biochi.2018.07.017;
RA   Sukumar N., Liu S., Li W., Mathews F.S., Mitra B., Kandavelu P.;
RT   "Structure of the monotopic membrane protein (S)-mandelate dehydrogenase at
RT   2.2A resolution.";
RL   Biochimie 154:45-54(2018).
CC   -!- FUNCTION: Catalyzes the dehydrogenation of (S)-mandelate to
CC       phenylglyoxylate (benzoylformate) (PubMed:10493804, PubMed:19465768).
CC       Is likely involved in the utilization of mandelate as a sole source of
CC       carbon and energy for growth (Probable). Active in vitro with the
CC       artificial electron acceptors 2,6-dichlorophenolindophenol (DCPIP) or
CC       ferricyanide, but in vivo most likely transfer the electron pair from
CC       the reduced flavin to a component of the electron transport chain in
CC       the membrane, possibly a quinone (PubMed:10493804, PubMed:19465768).
CC       Shows very low activity with oxygen as the electron acceptor, and also
CC       with 3-indolelactate and medium chain 2-hydroxyacids as substrates
CC       (PubMed:10493804, PubMed:19465768). {ECO:0000269|PubMed:10493804,
CC       ECO:0000269|PubMed:19465768, ECO:0000305|PubMed:2271624}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(S)-mandelate + A = phenylglyoxylate + AH2;
CC         Xref=Rhea:RHEA:15749, ChEBI:CHEBI:13193, ChEBI:CHEBI:17499,
CC         ChEBI:CHEBI:17756, ChEBI:CHEBI:36656; EC=1.1.99.31;
CC         Evidence={ECO:0000269|PubMed:10493804, ECO:0000269|PubMed:19465768};
CC       PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:15750;
CC         Evidence={ECO:0000305|PubMed:10493804};
CC   -!- COFACTOR:
CC       Name=FMN; Xref=ChEBI:CHEBI:58210;
CC         Evidence={ECO:0000269|PubMed:10493804, ECO:0000269|PubMed:11502180,
CC         ECO:0000269|PubMed:14604988, ECO:0000269|PubMed:19465768,
CC         ECO:0000269|PubMed:30071260};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=160 uM for (S)-mandelate (at pH 7.5 and 20 degrees Celsius)
CC         {ECO:0000269|PubMed:10493804};
CC         KM=750 uM for 2-hydroxyoctanoate (at pH 7.5 and 20 degrees Celsius)
CC         {ECO:0000269|PubMed:10493804};
CC         KM=4900 uM for 2-hydroxyhexanoate (at pH 7.5 and 20 degrees Celsius)
CC         {ECO:0000269|PubMed:10493804};
CC         KM=32000 uM for 2-hydroxybutanoate (at pH 7.5 and 20 degrees Celsius)
CC         {ECO:0000269|PubMed:10493804};
CC         KM=900 uM for 3-indolelactate (at pH 7.5 and 20 degrees Celsius)
CC         {ECO:0000269|PubMed:10493804, ECO:0000269|PubMed:19465768};
CC         KM=240 uM for (RS)-mandelate (at pH 7.5 and 20 degrees Celsius)
CC         {ECO:0000269|PubMed:19465768};
CC         KM=800 uM for 2-hydroxyoctanoate (at pH 7.5 and 20 degrees Celsius)
CC         {ECO:0000269|PubMed:19465768};
CC         Note=kcat is 290 sec(-1) with (S)-mandelate as substrate and DCPIP as
CC         electron acceptor (at pH 7.5 and 20 degrees Celsius)
CC         (PubMed:10493804). kcat is 0.5 sec(-1) with 2-hydroxyoctanoate as
CC         substrate and DCPIP as electron acceptor (at pH 7.5 and 20 degrees
CC         Celsius) (PubMed:10493804, PubMed:19465768). kcat is 0.34 sec(-1)
CC         with 2-hydroxyhexanoate as substrate and DCPIP as electron acceptor
CC         (at pH 7.5 and 20 degrees Celsius) (PubMed:10493804). kcat is 0.10
CC         sec(-1) with 2-hydroxybutanoate as substrate and DCPIP as electron
CC         acceptor (at pH 7.5 and 20 degrees Celsius) (PubMed:10493804). kcat
CC         is 1.0 sec(-1) with 3-indolelactate as substrate and DCPIP as
CC         electron acceptor (at pH 7.5 and 20 degrees Celsius)
CC         (PubMed:10493804, PubMed:19465768). kcat is 350 sec(-1) with
CC         (RS)-mandelate as substrate and DCPIP as electron acceptor (at pH 7.5
CC         and 20 degrees Celsius) (PubMed:19465768).
CC         {ECO:0000269|PubMed:10493804, ECO:0000269|PubMed:19465768};
CC   -!- PATHWAY: Aromatic compound metabolism; (R)-mandelate degradation;
CC       benzoate from (R)-mandelate: step 2/4. {ECO:0000305|PubMed:2271624}.
CC   -!- SUBUNIT: Homotetramer. {ECO:0000305|PubMed:11502180,
CC       ECO:0000305|PubMed:14604988}.
CC   -!- SUBCELLULAR LOCATION: Cell inner membrane
CC       {ECO:0000269|PubMed:10493804}.
CC   -!- SIMILARITY: Belongs to the FMN-dependent alpha-hydroxy acid
CC       dehydrogenase family. {ECO:0000255|PROSITE-ProRule:PRU00683}.
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DR   EMBL; AY143338; AAC15503.1; -; Genomic_DNA.
DR   PIR; B44767; B44767.
DR   PDB; 1HUV; X-ray; 2.15 A; A=1-176, A=216-393.
DR   PDB; 1P4C; X-ray; 1.35 A; A=1-176, A=216-393.
DR   PDB; 1P5B; X-ray; 1.35 A; A=1-176, A=216-393.
DR   PDB; 2A7N; X-ray; 1.80 A; A=1-176, A=216-393.
DR   PDB; 2A7P; X-ray; 2.20 A; A=1-176, A=216-393.
DR   PDB; 2A85; X-ray; 2.50 A; A=1-176, A=216-393.
DR   PDB; 3GIY; X-ray; 1.60 A; A=1-393.
DR   PDB; 6BFG; X-ray; 2.20 A; A/B=1-393.
DR   AlphaFoldDB; P20932; -.
DR   SMR; P20932; -.
DR   DrugBank; DB07907; (2S)-2-HYDROXYOCTANOIC ACID.
DR   DrugBank; DB03814; 2-(N-morpholino)ethanesulfonic acid.
DR   DrugBank; DB07060; 3-(INDOL-3-YL) LACTATE.
DR   DrugBank; DB03247; Flavin mononucleotide.
DR   BioCyc; MetaCyc:MONOMER-2422; -.
DR   BRENDA; 1.1.99.31; 5092.
DR   SABIO-RK; P20932; -.
DR   UniPathway; UPA00873; UER00853.
DR   EvolutionaryTrace; P20932; -.
DR   GO; GO:0005886; C:plasma membrane; EXP:UniProtKB.
DR   GO; GO:0033720; F:(S)-mandelate dehydrogenase activity; EXP:UniProtKB.
DR   GO; GO:0010181; F:FMN binding; IEA:InterPro.
DR   GO; GO:0004459; F:L-lactate dehydrogenase (NAD+) activity; IEA:TreeGrafter.
DR   GO; GO:0009060; P:aerobic respiration; IEA:TreeGrafter.
DR   GO; GO:0019596; P:mandelate catabolic process; IEA:UniProtKB-UniPathway.
DR   CDD; cd04736; MDH_FMN; 1.
DR   FunFam; 3.20.20.70:FF:000029; L-lactate dehydrogenase; 1.
DR   Gene3D; 3.20.20.70; Aldolase class I; 1.
DR   InterPro; IPR013785; Aldolase_TIM.
DR   InterPro; IPR012133; Alpha-hydoxy_acid_DH_FMN.
DR   InterPro; IPR000262; FMN-dep_DH.
DR   InterPro; IPR037396; FMN_HAD.
DR   InterPro; IPR008259; FMN_hydac_DH_AS.
DR   PANTHER; PTHR10578:SF107; 2-HYDROXYACID OXIDASE 1; 1.
DR   PANTHER; PTHR10578; S -2-HYDROXY-ACID OXIDASE-RELATED; 1.
DR   Pfam; PF01070; FMN_dh; 1.
DR   PIRSF; PIRSF000138; Al-hdrx_acd_dh; 1.
DR   SUPFAM; SSF51395; FMN-linked oxidoreductases; 1.
DR   PROSITE; PS00557; FMN_HYDROXY_ACID_DH_1; 1.
DR   PROSITE; PS51349; FMN_HYDROXY_ACID_DH_2; 1.
DR   PDBsum; 1HUV; -.
DR   PDBsum; 1P4C; -.
DR   PDBsum; 1P5B; -.
DR   PDBsum; 2A7N; -.
DR   PDBsum; 2A7P; -.
DR   PDBsum; 2A85; -.
DR   PDBsum; 3GIY; -.
DR   PDBsum; 6BFG; -.
PE   1: Evidence at protein level;
KW   3D-structure; Aromatic hydrocarbons catabolism; Cell inner membrane;
KW   Cell membrane; Direct protein sequencing; Flavoprotein; FMN;
KW   Mandelate pathway; Membrane; Oxidoreductase.
FT   CHAIN           1..393
FT                   /note="(S)-mandelate dehydrogenase"
FT                   /id="PRO_0000206327"
FT   DOMAIN          1..377
FT                   /note="FMN hydroxy acid dehydrogenase"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00683"
FT   ACT_SITE        274
FT                   /note="Proton acceptor"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00683"
FT   BINDING         26
FT                   /ligand="(S)-mandelate"
FT                   /ligand_id="ChEBI:CHEBI:17756"
FT                   /evidence="ECO:0000305|PubMed:19465768,
FT                   ECO:0007744|PDB:2A85"
FT   BINDING         79..81
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000269|PubMed:14604988,
FT                   ECO:0000269|PubMed:30071260, ECO:0007744|PDB:1P4C,
FT                   ECO:0007744|PDB:6BFG"
FT   BINDING         108
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000269|PubMed:14604988,
FT                   ECO:0000269|PubMed:30071260, ECO:0007744|PDB:1P4C,
FT                   ECO:0007744|PDB:6BFG"
FT   BINDING         129
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000269|PubMed:14604988,
FT                   ECO:0000269|PubMed:30071260, ECO:0007744|PDB:1P4C,
FT                   ECO:0007744|PDB:6BFG"
FT   BINDING         131
FT                   /ligand="(S)-mandelate"
FT                   /ligand_id="ChEBI:CHEBI:17756"
FT                   /evidence="ECO:0000305|PubMed:19465768,
FT                   ECO:0007744|PDB:2A85"
FT   BINDING         156
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000269|PubMed:14604988,
FT                   ECO:0000269|PubMed:30071260, ECO:0007744|PDB:1P4C,
FT                   ECO:0007744|PDB:6BFG"
FT   BINDING         165
FT                   /ligand="(S)-mandelate"
FT                   /ligand_id="ChEBI:CHEBI:17756"
FT                   /evidence="ECO:0000305|PubMed:19465768,
FT                   ECO:0007744|PDB:2A85"
FT   BINDING         250
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000269|PubMed:14604988,
FT                   ECO:0000269|PubMed:30071260, ECO:0007744|PDB:1P4C,
FT                   ECO:0007744|PDB:6BFG"
FT   BINDING         274
FT                   /ligand="(S)-mandelate"
FT                   /ligand_id="ChEBI:CHEBI:17756"
FT                   /evidence="ECO:0000305|PubMed:19465768,
FT                   ECO:0007744|PDB:2A85"
FT   BINDING         277
FT                   /ligand="(S)-mandelate"
FT                   /ligand_id="ChEBI:CHEBI:17756"
FT                   /evidence="ECO:0000305|PubMed:19465768,
FT                   ECO:0007744|PDB:2A85"
FT   BINDING         303..307
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000269|PubMed:14604988,
FT                   ECO:0000269|PubMed:30071260, ECO:0007744|PDB:1P4C,
FT                   ECO:0007744|PDB:6BFG"
FT   BINDING         326..327
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000269|PubMed:14604988,
FT                   ECO:0000269|PubMed:30071260, ECO:0007744|PDB:1P4C,
FT                   ECO:0007744|PDB:6BFG"
FT   MUTAGEN         81
FT                   /note="G->A: 23-fold decrease in catalytic activity with
FT                   mandelate as substrate and DCPIP as electron acceptor, but
FT                   no change in affinity for mandelate. Shows a modestly
FT                   higher reactivity with molecular oxygen."
FT                   /evidence="ECO:0000269|PubMed:19465768"
FT   HELIX           8..14
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           16..18
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           21..28
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           35..42
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           43..46
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          47..49
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           51..53
FT                   /evidence="ECO:0007829|PDB:6BFG"
FT   STRAND          64..66
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          69..77
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           83..85
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           90..101
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          105..107
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           115..121
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          126..130
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           135..147
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          152..156
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           166..171
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           181..188
FT                   /evidence="ECO:0007829|PDB:6BFG"
FT   HELIX           191..200
FT                   /evidence="ECO:0007829|PDB:6BFG"
FT   HELIX           206..208
FT                   /evidence="ECO:0007829|PDB:6BFG"
FT   STRAND          210..213
FT                   /evidence="ECO:0007829|PDB:6BFG"
FT   TURN            216..218
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           219..222
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           233..242
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          245..252
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           255..263
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          267..271
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           274..276
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           282..284
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           285..287
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           289..296
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          300..302
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           309..317
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          323..326
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           327..359
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           364..366
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   HELIX           369..371
FT                   /evidence="ECO:0007829|PDB:1P4C"
FT   STRAND          372..374
FT                   /evidence="ECO:0007829|PDB:1P4C"
SQ   SEQUENCE   393 AA;  43437 MW;  18BE23E459BB3987 CRC64;
     MSQNLFNVED YRKLRQKRLP KMVYDYLEGG AEDEYGVKHN RDVFQQWRFK PKRLVDVSRR
     SLQAEVLGKR QSMPLLIGPT GLNGALWPKG DLALARAATK AGIPFVLSTA SNMSIEDLAR
     QCDGDLWFQL YVIHREIAQG MVLKALHTGY TTLVLTTDVA VNGYRERDLH NRFKIPMSYS
     AKVVLDGCLH PRWSLDFVRH GMPQLANFVS SQTSSLEMQA ALMSRQMDAS FNWEALRWLR
     DLWPHKLLVK GLLSAEDADR CIAEGADGVI LSNHGGRQLD CAISPMEVLA QSVAKTGKPV
     LIDSGFRRGS DIVKALALGA EAVLLGRATL YGLAARGETG VDEVLTLLKA DIDRTLAQIG
     CPDITSLSPD YLQNEGVTNT APVDHLIGKG THA
//