MDLB_PSEPU
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 03-AUG-2022, entry version 118.
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; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC 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 = AH2 + phenylglyoxylate;
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 (RS)-
CC mandelate as substrate and DCPIP as electron acceptor (at pH 7.5 and
CC 20 degrees Celsius) (PubMed:19465768). {ECO:0000269|PubMed:10493804,
CC 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=1-176, A=216-393.
DR PDB; 6BFG; X-ray; 2.20 A; A/B=1-393.
DR PDBsum; 1HUV; -.
DR PDBsum; 1P4C; -.
DR PDBsum; 1P5B; -.
DR PDBsum; 2A7N; -.
DR PDBsum; 2A7P; -.
DR PDBsum; 2A85; -.
DR PDBsum; 3GIY; -.
DR PDBsum; 6BFG; -.
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:MON-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; IEA:UniProtKB-SubCell.
DR GO; GO:0033720; F:(S)-mandelate dehydrogenase activity; IEA:UniProtKB-EC.
DR GO; GO:0010181; F:FMN binding; IEA:InterPro.
DR GO; GO:0019596; P:mandelate catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.20.20.70; -; 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 Pfam; PF01070; FMN_dh; 1.
DR PIRSF; PIRSF000138; Al-hdrx_acd_dh; 1.
DR PROSITE; PS00557; FMN_HYDROXY_ACID_DH_1; 1.
DR PROSITE; PS51349; FMN_HYDROXY_ACID_DH_2; 1.
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 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 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
