ID   FCL_ECOLI               Reviewed;         321 AA.
AC   P32055; P76382;
DT   01-OCT-1993, integrated into UniProtKB/Swiss-Prot.
DT   01-NOV-1997, sequence version 2.
DT   02-SEP-2026, entry version 187.
DE   RecName: Full=GDP-L-fucose synthase {ECO:0000255|HAMAP-Rule:MF_00956};
DE            EC=1.1.1.271 {ECO:0000255|HAMAP-Rule:MF_00956, ECO:0000269|PubMed:10480878, ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9473059};
DE   AltName: Full=GDP-4-keto-6-deoxy-D-mannose-3,5-epimerase-4-reductase {ECO:0000255|HAMAP-Rule:MF_00956};
GN   Name=fcl {ECO:0000255|HAMAP-Rule:MF_00956}; Synonyms=wcaG, yefB;
GN   OrderedLocusNames=b2052, JW2037;
OS   Escherichia coli (strain K12).
OC   Bacteria; Pseudomonadati; Pseudomonadota; Gammaproteobacteria;
OC   Enterobacterales; Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=83333;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=K12;
RX   PubMed=7815923; DOI=10.1093/oxfordjournals.molbev.a040166;
RA   Aoyama K., Haase A.M., Reeves P.R.;
RT   "Evidence for effect of random genetic drift on G+C content after lateral
RT   transfer of fucose pathway genes to Escherichia coli K-12.";
RL   Mol. Biol. Evol. 11:829-838(1994).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=K12;
RX   PubMed=8759852; DOI=10.1128/jb.178.16.4885-4893.1996;
RA   Stevenson G., Andrianopoulos K., Hobbs M., Reeves P.R.;
RT   "Organization of the Escherichia coli K-12 gene cluster responsible for
RT   production of the extracellular polysaccharide colanic acid.";
RL   J. Bacteriol. 178:4885-4893(1996).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=9097040; DOI=10.1093/dnares/3.6.379;
RA   Itoh T., Aiba H., Baba T., Fujita K., Hayashi K., Inada T., Isono K.,
RA   Kasai H., Kimura S., Kitakawa M., Kitagawa M., Makino K., Miki T.,
RA   Mizobuchi K., Mori H., Mori T., Motomura K., Nakade S., Nakamura Y.,
RA   Nashimoto H., Nishio Y., Oshima T., Saito N., Sampei G., Seki Y.,
RA   Sivasundaram S., Tagami H., Takeda J., Takemoto K., Wada C., Yamamoto Y.,
RA   Horiuchi T.;
RT   "A 460-kb DNA sequence of the Escherichia coli K-12 genome corresponding to
RT   the 40.1-50.0 min region on the linkage map.";
RL   DNA Res. 3:379-392(1996).
RN   [4]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA   Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA   Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA   Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA   Shao Y.;
RT   "The complete genome sequence of Escherichia coli K-12.";
RL   Science 277:1453-1462(1997).
RN   [5]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=16738553; DOI=10.1038/msb4100049;
RA   Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA   Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT   "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT   and W3110.";
RL   Mol. Syst. Biol. 2:E1-E5(2006).
RN   [6]
RP   FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX   PubMed=9473059; DOI=10.1128/jb.180.4.998-1001.1998;
RA   Andrianopoulos K., Wang L., Reeves P.R.;
RT   "Identification of the fucose synthetase gene in the colanic acid gene
RT   cluster of Escherichia coli K-12.";
RL   J. Bacteriol. 180:998-1001(1998).
RN   [7]
RP   FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL
RP   PROPERTIES.
RX   PubMed=10480878; DOI=10.1074/jbc.274.38.26743;
RA   Menon S., Stahl M., Kumar R., Xu G.Y., Sullivan F.;
RT   "Stereochemical course and steady state mechanism of the reaction catalyzed
RT   by the GDP-fucose synthetase from Escherichia coli.";
RL   J. Biol. Chem. 274:26743-26750(1999).
RN   [8]
RP   X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH NADP, AND SUBUNIT.
RX   PubMed=9817848; DOI=10.1016/s0969-2126(98)00144-0;
RA   Rizzi M., Tonetti M., Vigevani P., Sturla L., Bisso A., Flora A.D.,
RA   Bordo D., Bolognesi M.;
RT   "GDP-4-keto-6-deoxy-D-mannose epimerase/reductase from Escherichia coli, a
RT   key enzyme in the biosynthesis of GDP-L-fucose, displays the structural
RT   characteristics of the RED protein homology superfamily.";
RL   Structure 6:1453-1465(1998).
RN   [9]
RP   X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH NADP.
RC   STRAIN=K12;
RX   PubMed=9862812; DOI=10.1016/s0969-2126(98)00157-9;
RA   Somers W.S., Stahl M.L., Sullivan F.X.;
RT   "GDP-fucose synthetase from Escherichia coli: structure of a unique member
RT   of the short-chain dehydrogenase/reductase family that catalyzes two
RT   distinct reactions at the same active site.";
RL   Structure 6:1601-1612(1998).
RN   [10]
RP   X-RAY CRYSTALLOGRAPHY (1.45 ANGSTROMS) IN COMPLEX WITH NADP, CATALYTIC
RP   ACTIVITY, FUNCTION, PREDICTED SUBSTRATE-BINDING SITES, MUTAGENESIS OF
RP   SER-107; TYR-136; LYS-140; HIS-179 AND ARG-187, BIOPHYSICOCHEMICAL
RP   PROPERTIES, AND ACTIVE SITE.
RX   PubMed=11021971; DOI=10.1006/jmbi.2000.4106;
RA   Rosano C., Bisso A., Izzo G., Tonetti M., Sturla L., De Flora A.,
RA   Bolognesi M.;
RT   "Probing the catalytic mechanism of GDP-4-keto-6-deoxy-d-mannose
RT   Epimerase/Reductase by kinetic and crystallographic characterization of
RT   site-specific mutants.";
RL   J. Mol. Biol. 303:77-91(2000).
CC   -!- FUNCTION: Catalyzes the two-step NADP-dependent conversion of GDP-4-
CC       dehydro-6-deoxy-D-mannose to GDP-fucose, involving an epimerase and a
CC       reductase reaction. {ECO:0000255|HAMAP-Rule:MF_00956,
CC       ECO:0000269|PubMed:10480878, ECO:0000269|PubMed:11021971,
CC       ECO:0000269|PubMed:9473059}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=GDP-beta-L-fucose + NADP(+) = GDP-4-dehydro-alpha-D-rhamnose +
CC         NADPH + H(+); Xref=Rhea:RHEA:18885, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:57273, ChEBI:CHEBI:57783, ChEBI:CHEBI:57964,
CC         ChEBI:CHEBI:58349; EC=1.1.1.271; Evidence={ECO:0000255|HAMAP-
CC         Rule:MF_00956, ECO:0000269|PubMed:10480878,
CC         ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9473059};
CC   -!- ACTIVITY REGULATION: Subject to product inhibition by NADP and GDP-
CC       fucose. {ECO:0000269|PubMed:10480878}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=9 uM for NADPH {ECO:0000269|PubMed:10480878,
CC         ECO:0000269|PubMed:11021971};
CC         KM=109 uM for NADH {ECO:0000269|PubMed:10480878,
CC         ECO:0000269|PubMed:11021971};
CC         KM=29 uM for GDP-4-keto-6-deoxymannose {ECO:0000269|PubMed:10480878,
CC         ECO:0000269|PubMed:11021971};
CC         Vmax=363.5 umol/min/mg enzyme {ECO:0000269|PubMed:10480878,
CC         ECO:0000269|PubMed:11021971};
CC       pH dependence:
CC         Optimum pH is 6-6.5. {ECO:0000269|PubMed:10480878,
CC         ECO:0000269|PubMed:11021971};
CC   -!- PATHWAY: Nucleotide-sugar biosynthesis; GDP-L-fucose biosynthesis via
CC       de novo pathway; GDP-L-fucose from GDP-alpha-D-mannose: step 2/2.
CC       {ECO:0000255|HAMAP-Rule:MF_00956, ECO:0000269|PubMed:9473059}.
CC   -!- PATHWAY: Exopolysaccharide biosynthesis; colanic acid biosynthesis.
CC       {ECO:0000269|PubMed:9473059}.
CC   -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:11021971,
CC       ECO:0000269|PubMed:9817848, ECO:0000269|PubMed:9862812}.
CC   -!- SUBCELLULAR LOCATION: Cytoplasm.
CC   -!- SIMILARITY: Belongs to the NAD(P)-dependent epimerase/dehydratase
CC       family. Fucose synthase subfamily. {ECO:0000255|HAMAP-Rule:MF_00956}.
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DR   EMBL; U38473; AAC77843.1; -; Genomic_DNA.
DR   EMBL; U00096; AAC75113.1; -; Genomic_DNA.
DR   EMBL; AP009048; BAA15908.1; -; Genomic_DNA.
DR   PIR; C64971; C64971.
DR   RefSeq; NP_416556.1; NC_000913.3.
DR   RefSeq; WP_000043654.1; NZ_CP009789.1.
DR   PDB; 1BSV; X-ray; 2.20 A; A=1-321.
DR   PDB; 1BWS; X-ray; 2.20 A; A=1-321.
DR   PDB; 1E6U; X-ray; 1.45 A; A=1-321.
DR   PDB; 1E7Q; X-ray; 1.60 A; A=1-321.
DR   PDB; 1E7R; X-ray; 1.60 A; A=1-321.
DR   PDB; 1E7S; X-ray; 1.50 A; A=1-321.
DR   PDB; 1FXS; X-ray; 2.30 A; A=1-321.
DR   PDB; 1GFS; X-ray; 2.20 A; A=1-321.
DR   AlphaFoldDB; P32055; -.
DR   SMR; P32055; -.
DR   BioGRID; 4259691; 222.
DR   FunCoup; P32055; 420.
DR   IntAct; P32055; 2.
DR   STRING; 511145.b2052; -.
DR   DrugBank; DB02897; Acetylphosphate.
DR   DrugBank; DB03461; Nicotinamide adenine dinucleotide phosphate.
DR   PaxDb; 511145-b2052; -.
DR   EnsemblBacteria; AAC75113; AAC75113; b2052.
DR   GeneID; 946563; -.
DR   KEGG; ecj:JW2037; -.
DR   KEGG; eco:b2052; -.
DR   KEGG; ecoc:C3026_11550; -.
DR   PATRIC; fig|1411691.4.peg.199; -.
DR   EchoBASE; EB1736; -.
DR   eggNOG; COG0451; Bacteria.
DR   HOGENOM; CLU_007383_18_0_6; -.
DR   InParanoid; P32055; -.
DR   OMA; HPSNYGY; -.
DR   OrthoDB; 9811425at2; -.
DR   PhylomeDB; P32055; -.
DR   BioCyc; EcoCyc:FCL-MONOMER; -.
DR   BioCyc; MetaCyc:FCL-MONOMER; -.
DR   BRENDA; 1.1.1.271; 2026.
DR   SABIO-RK; P32055; -.
DR   UniPathway; UPA00128; UER00191.
DR   UniPathway; UPA00980; -.
DR   EvolutionaryTrace; P32055; -.
DR   PRO; PR:P32055; -.
DR   Proteomes; UP000000625; Chromosome.
DR   GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR   GO; GO:0050577; F:GDP-L-fucose synthase activity; IDA:EcoCyc.
DR   GO; GO:0070401; F:NADP+ binding; IEA:UniProtKB-UniRule.
DR   GO; GO:0042803; F:protein homodimerization activity; IDA:EcoCyc.
DR   GO; GO:0042351; P:'de novo' GDP-L-fucose biosynthetic process; IEA:UniProtKB-UniRule.
DR   GO; GO:0009242; P:colanic acid biosynthetic process; IEA:UniProtKB-UniPathway.
DR   CDD; cd05239; GDP_FS_SDR_e; 1.
DR   FunFam; 3.40.50.720:FF:000101; GDP-L-fucose synthase; 1.
DR   Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1.
DR   Gene3D; 3.90.25.10; UDP-galactose 4-epimerase, domain 1; 1.
DR   HAMAP; MF_00956; GDP_fucose_synth; 1.
DR   InterPro; IPR001509; Epimerase_deHydtase.
DR   InterPro; IPR028614; GDP_fucose/colitose_synth.
DR   InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR   NCBIfam; NF047922; GDPFucSynFcl; 1.
DR   PANTHER; PTHR43238; GDP-L-FUCOSE SYNTHASE; 1.
DR   PANTHER; PTHR43238:SF1; GDP-L-FUCOSE SYNTHASE; 1.
DR   Pfam; PF01370; Epimerase; 1.
DR   SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1.
DR   PDBsum; 1BSV; -.
DR   PDBsum; 1BWS; -.
DR   PDBsum; 1E6U; -.
DR   PDBsum; 1E7Q; -.
DR   PDBsum; 1E7R; -.
DR   PDBsum; 1E7S; -.
DR   PDBsum; 1FXS; -.
DR   PDBsum; 1GFS; -.
PE   1: Evidence at protein level;
KW   3D-structure; Cytoplasm; Isomerase; Multifunctional enzyme; NADP;
KW   Oxidoreductase; Reference proteome.
FT   CHAIN           1..321
FT                   /note="GDP-L-fucose synthase"
FT                   /id="PRO_0000174358"
FT   ACT_SITE        136
FT                   /note="Proton donor/acceptor"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT                   ECO:0000269|PubMed:11021971"
FT   BINDING         10..16
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT                   ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT                   ECO:0000269|PubMed:9862812"
FT   BINDING         36..41
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT                   ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT                   ECO:0000269|PubMed:9862812"
FT   BINDING         105..108
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT                   ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT                   ECO:0000269|PubMed:9862812"
FT   BINDING         140
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT                   ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT                   ECO:0000269|PubMed:9862812"
FT   BINDING         163..166
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT                   ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT                   ECO:0000269|PubMed:9862812"
FT   BINDING         179
FT                   /ligand="NADP(+)"
FT                   /ligand_id="ChEBI:CHEBI:58349"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956,
FT                   ECO:0000269|PubMed:11021971, ECO:0000269|PubMed:9817848,
FT                   ECO:0000269|PubMed:9862812"
FT   BINDING         187
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000305"
FT   BINDING         202
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956"
FT   BINDING         209
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956"
FT   BINDING         278
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_00956"
FT   SITE            107
FT                   /note="Important for catalytic activity"
FT   SITE            109
FT                   /note="Important for catalytic activity"
FT   SITE            140
FT                   /note="Lowers pKa of active site Tyr"
FT   MUTAGEN         107
FT                   /note="S->A: Nearly abolishes catalytic activity. Minor
FT                   effect of affinity for NADPH and substrate."
FT                   /evidence="ECO:0000269|PubMed:11021971"
FT   MUTAGEN         109
FT                   /note="C->A: Nearly abolishes catalytic activity."
FT   MUTAGEN         136
FT                   /note="Y->E: Abolishes enzyme activity."
FT                   /evidence="ECO:0000269|PubMed:11021971"
FT   MUTAGEN         140
FT                   /note="K->R: Reduces catalytic activity 20-fold."
FT                   /evidence="ECO:0000269|PubMed:11021971"
FT   MUTAGEN         140
FT                   /note="K->S: Nearly abolishes catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:11021971"
FT   MUTAGEN         179
FT                   /note="H->N: Nearly abolishes catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:11021971"
FT   MUTAGEN         187
FT                   /note="R->A: Decreases affinity for the substrate
FT                   GDP-4-keto-6-deoxymannose."
FT                   /evidence="ECO:0000269|PubMed:11021971"
FT   CONFLICT        255..256
FT                   /note="EL -> DV (in Ref. 1; no nucleotide entry and 2;
FT                   AAC77843)"
FT                   /evidence="ECO:0000305"
FT   STRAND          4..9
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   TURN            10..12
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           14..23
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          29..32
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   TURN            36..38
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           44..54
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          57..61
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           69..74
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           76..96
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          101..105
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           108..110
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          117..119
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           121..123
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           128..130
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           131..133
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           134..154
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          157..163
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          165..168
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           180..194
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          197..203
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          211..213
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           214..226
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           229..234
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          243..246
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           253..264
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          268..273
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   STRAND          281..283
FT                   /evidence="ECO:0007829|PDB:1BSV"
FT   HELIX           288..292
FT                   /evidence="ECO:0007829|PDB:1E6U"
FT   HELIX           301..314
FT                   /evidence="ECO:0007829|PDB:1E6U"
SQ   SEQUENCE   321 AA;  36141 MW;  97077193D79684C7 CRC64;
     MSKQRVFIAG HRGMVGSAIR RQLEQRGDVE LVLRTRDELN LLDSRAVHDF FASERIDQVY
     LAAAKVGGIV ANNTYPADFI YQNMMIESNI IHAAHQNDVN KLLFLGSSCI YPKLAKQPMA
     ESELLQGTLE PTNEPYAIAK IAGIKLCESY NRQYGRDYRS VMPTNLYGPH DNFHPSNSHV
     IPALLRRFHE ATAQNAPDVV VWGSGTPMRE FLHVDDMAAA SIHVMELAHE VWLENTQPML
     SHINVGTGVD CTIRELAQTI AKVVGYKGRV VFDASKPDGT PRKLLDVTRL HQLGWYHEIS
     LEAGLASTYQ WFLENQDRFR G
//