ID ULAD_ECOLI Reviewed; 216 AA. AC P39304; Q2M6A9; DT 01-FEB-1995, integrated into UniProtKB/Swiss-Prot. DT 01-FEB-1995, sequence version 1. DT 24-JAN-2024, entry version 165. DE RecName: Full=3-keto-L-gulonate-6-phosphate decarboxylase UlaD; DE EC=4.1.1.85 {ECO:0000269|PubMed:11741871}; DE AltName: Full=3-dehydro-L-gulonate-6-phosphate decarboxylase; DE AltName: Full=KGPDC; DE AltName: Full=L-ascorbate utilization protein D; GN Name=ulaD; Synonyms=sgaH, yjfV; OrderedLocusNames=b4196, JW4154; OS Escherichia coli (strain K12). OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales; OC Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=7610040; DOI=10.1093/nar/23.12.2105; RA Burland V.D., Plunkett G. III, Sofia H.J., Daniels D.L., Blattner F.R.; RT "Analysis of the Escherichia coli genome VI: DNA sequence of the region RT from 92.8 through 100 minutes."; RL Nucleic Acids Res. 23:2105-2119(1995). RN [2] 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 [3] 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 [4] RP DISCUSSION OF SEQUENCE. RA Reizer J., Charbit A., Reizer A., Saier M.H. Jr.; RT "Novel phosphotransferases system genes revealed by bacterial genome RT analysis: operons encoding homologues of sugar-specific permease domains of RT the phosphotransferase system and pentose catabolic enzymes."; RL Genome Sci. Technol. 1:53-75(1996). RN [5] RP DISCUSSION OF SEQUENCE. RX PubMed=9274005; DOI=10.1099/00221287-143-8-2519; RA Reizer J., Reizer A., Saier M.H. Jr.; RT "Is the ribulose monophosphate pathway widely distributed in bacteria?"; RL Microbiology 143:2519-2520(1997). RN [6] RP FUNCTION, CATALYTIC ACTIVITY, AND ROLE IN L-ASCORBATE UTILIZATION. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=11741871; DOI=10.1128/jb.184.1.302-306.2002; RA Yew W.S., Gerlt J.A.; RT "Utilization of L-ascorbate by Escherichia coli K-12: assignments of RT functions to products of the yjf-sga and yia-sgb operons."; RL J. Bacteriol. 184:302-306(2002). RN [7] RP TRANSCRIPTIONAL REGULATION. RX PubMed=12374842; DOI=10.1128/jb.184.21.6065-6068.2002; RA Campos E., Aguilar J., Baldoma L., Badia J.; RT "The gene yjfQ encodes the repressor of the yjfR-X regulon (ula), which is RT involved in L-ascorbate metabolism in Escherichia coli."; RL J. Bacteriol. 184:6065-6068(2002). RN [8] RP TRANSCRIPTIONAL REGULATION. RX PubMed=14996803; DOI=10.1128/jb.186.6.1720-1728.2004; RA Campos E., Baldoma L., Aguilar J., Badia J.; RT "Regulation of expression of the divergent ulaG and ulaABCDEF operons RT involved in L-ascorbate dissimilation in Escherichia coli."; RL J. Bacteriol. 186:1720-1728(2004). RN [9] RP MUTAGENESIS OF GLU-33; LYS-64; ASP-67; GLU-112; HIS-136 AND ARG-139, RP KINETIC PARAMETERS, AND REACTION STEREOCHEMISTRY. RX PubMed=15157077; DOI=10.1021/bi049741t; RA Yew W.S., Wise E.L., Rayment I., Gerlt J.A.; RT "Evolution of enzymatic activities in the orotidine 5'-monophosphate RT decarboxylase suprafamily: mechanistic evidence for a proton relay system RT in the active site of 3-keto-L-gulonate 6-phosphate decarboxylase."; RL Biochemistry 43:6427-6437(2004). RN [10] RP MUTAGENESIS. RX PubMed=15697206; DOI=10.1021/bi047815v; RA Yew W.S., Akana J., Wise E.L., Rayment I., Gerlt J.A.; RT "Evolution of enzymatic activities in the orotidine 5'-monophosphate RT decarboxylase suprafamily: enhancing the promiscuous D-arabino-hex-3-ulose RT 6-phosphate synthase reaction catalyzed by 3-keto-L-gulonate 6-phosphate RT decarboxylase."; RL Biochemistry 44:1807-1815(2005). RN [11] RP X-RAY CRYSTALLOGRAPHY (1.62 ANGSTROMS) OF COMPLEXES WITH MAGNESIUM; RP PHOSPHATE AND L-GULONATE-6-PHOSPHATE, AND SUBUNIT. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=11900527; DOI=10.1021/bi012174e; RA Wise E., Yew W.S., Babbitt P.C., Gerlt J.A., Rayment I.; RT "Homologous (beta/alpha)8-barrel enzymes that catalyze unrelated reactions: RT orotidine 5'-monophosphate decarboxylase and 3-keto-L-gulonate 6-phosphate RT decarboxylase."; RL Biochemistry 41:3861-3869(2002). RN [12] RP X-RAY CRYSTALLOGRAPHY (1.2 ANGSTROMS) OF COMPLEXES WITH L-GULONATE RP 6-PHOSPHATE; L-THREONOHYDROXAMATE 4-PHOSPHATE; L-XYLITOL 5-PHOSPHATE AND RP L-XYLULOSE 5-PHOSPHATE. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=14567674; DOI=10.1021/bi0348819; RA Wise E.L., Yew W.S., Gerlt J.A., Rayment I.; RT "Structural evidence for a 1,2-enediolate intermediate in the reaction RT catalyzed by 3-keto-L-gulonate 6-phosphate decarboxylase, a member of the RT orotidine 5'-monophosphate decarboxylase suprafamily."; RL Biochemistry 42:12133-12142(2003). RN [13] RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) OF MUTANTS ALA-64; ALA-136; GLN-112 RP AND GLN-112/ALA-136 IN COMPLEX WITH L-THREONOHYDROXAMATE 4-PHOSPHATE, AND RP REACTION MECHANISM. RX PubMed=15157078; DOI=10.1021/bi0497392; RA Wise E.L., Yew W.S., Gerlt J.A., Rayment I.; RT "Evolution of enzymatic activities in the orotidine 5'-monophosphate RT decarboxylase suprafamily: crystallographic evidence for a proton relay RT system in the active site of 3-keto-L-gulonate 6-phosphate decarboxylase."; RL Biochemistry 43:6438-6446(2004). RN [14] RP X-RAY CRYSTALLOGRAPHY (1.6 ANGSTROMS) OF NATIVE PROTEIN IN COMPLEX WITH RP D-RIBULOSE 5-PHOSPHATE; MUTANTS ASP-112/ALA-169 AND ASP-11/VAL-139/ALA-169 RP IN COMPLEX WITH D-RIBULOSE 5-PHOSPHATE AND MUTANT ASP-11/VAL-139/ALA-169 IN RP COMPLEX WITH L-XYLULOSE 5-PHOSPHATE. RX PubMed=15697207; DOI=10.1021/bi0478143; RA Wise E.L., Yew W.S., Akana J., Gerlt J.A., Rayment I.; RT "Evolution of enzymatic activities in the orotidine 5'-monophosphate RT decarboxylase suprafamily: structural basis for catalytic promiscuity in RT wild-type and designed mutants of 3-keto-L-gulonate 6-phosphate RT decarboxylase."; RL Biochemistry 44:1816-1823(2005). CC -!- FUNCTION: Catalyzes the decarboxylation of 3-keto-L-gulonate-6-P into CC L-xylulose-5-P. Is involved in the anaerobic L-ascorbate utilization. CC {ECO:0000269|PubMed:11741871}. CC -!- CATALYTIC ACTIVITY: CC Reaction=3-dehydro-L-gulonate 6-phosphate + H(+) = CO2 + L-xylulose 5- CC phosphate; Xref=Rhea:RHEA:14353, ChEBI:CHEBI:15378, CC ChEBI:CHEBI:16526, ChEBI:CHEBI:57829, ChEBI:CHEBI:58774; EC=4.1.1.85; CC Evidence={ECO:0000269|PubMed:11741871}; CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:14354; CC Evidence={ECO:0000269|PubMed:11741871}; CC -!- COFACTOR: CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; CC Note=Binds 1 Mg(2+) ion per subunit.; CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=0.67 mM for 3-keto-L-gulonate-6-P {ECO:0000269|PubMed:15157077}; CC -!- PATHWAY: Cofactor degradation; L-ascorbate degradation; D-xylulose 5- CC phosphate from L-ascorbate: step 2/4. CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:11900527, CC ECO:0000269|PubMed:15157078, ECO:0000269|PubMed:15697207}. CC -!- INDUCTION: Induced by L-ascorbate. Repressed by UlaR. CC {ECO:0000269|PubMed:12374842, ECO:0000269|PubMed:14996803}. CC -!- MISCELLANEOUS: The reaction mechanism proceeds via the formation of a CC Mg(2+) ion-stabilized 1,2-cis-enediolate intermediate. Water molecules CC competitively shuttle protons from the side chains of His-136 and Arg- CC 139 to alternate faces of this intermediate. The active site is located CC at the interface of the component polypeptides. CC -!- SIMILARITY: Belongs to the HPS/KGPDC family. KGPDC subfamily. CC {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; U14003; AAA97092.1; -; Genomic_DNA. DR EMBL; U00096; AAC77153.1; -; Genomic_DNA. DR EMBL; AP009048; BAE78197.1; -; Genomic_DNA. DR PIR; S56421; S56421. DR RefSeq; NP_418617.1; NC_000913.3. DR RefSeq; WP_000056749.1; NZ_LN832404.1. DR PDB; 1KV8; X-ray; 1.62 A; A/B=1-216. DR PDB; 1KW1; X-ray; 2.20 A; A/B=1-216. DR PDB; 1Q6L; X-ray; 1.80 A; A/B=1-216. DR PDB; 1Q6O; X-ray; 1.20 A; A/B=1-216. DR PDB; 1Q6Q; X-ray; 1.70 A; A/B=1-216. DR PDB; 1Q6R; X-ray; 1.76 A; A/B=1-216. DR PDB; 1SO3; X-ray; 1.90 A; A/B=1-216. DR PDB; 1SO4; X-ray; 1.70 A; A/B=1-216. DR PDB; 1SO5; X-ray; 1.80 A; A/B=1-216. DR PDB; 1SO6; X-ray; 1.90 A; A/B=1-216. DR PDB; 1XBV; X-ray; 1.66 A; A/B=1-216. DR PDB; 1XBX; X-ray; 1.81 A; A/B=1-216. DR PDB; 1XBY; X-ray; 1.58 A; A/B=1-216. DR PDB; 1XBZ; X-ray; 1.80 A; A/B=1-216. DR PDBsum; 1KV8; -. DR PDBsum; 1KW1; -. DR PDBsum; 1Q6L; -. DR PDBsum; 1Q6O; -. DR PDBsum; 1Q6Q; -. DR PDBsum; 1Q6R; -. DR PDBsum; 1SO3; -. DR PDBsum; 1SO4; -. DR PDBsum; 1SO5; -. DR PDBsum; 1SO6; -. DR PDBsum; 1XBV; -. DR PDBsum; 1XBX; -. DR PDBsum; 1XBY; -. DR PDBsum; 1XBZ; -. DR AlphaFoldDB; P39304; -. DR SMR; P39304; -. DR BioGRID; 4262000; 15. DR DIP; DIP-10869N; -. DR IntAct; P39304; 1. DR STRING; 511145.b4196; -. DR DrugBank; DB01655; L-gulonic acid 6-phosphate. DR DrugBank; DB03855; L-Threonohydroxamate 4-Phosphate. DR DrugBank; DB02630; L-xylitol 5-phosphate. DR DrugBank; DB01923; L-Xylulose 5-Phosphate. DR DrugBank; DB04034; Ribulose-5-Phosphate. DR PaxDb; 511145-b4196; -. DR EnsemblBacteria; AAC77153; AAC77153; b4196. DR GeneID; 948714; -. DR KEGG; ecj:JW4154; -. DR KEGG; eco:b4196; -. DR PATRIC; fig|1411691.4.peg.2505; -. DR EchoBASE; EB2389; -. DR eggNOG; COG0269; Bacteria. DR HOGENOM; CLU_081825_0_0_6; -. DR InParanoid; P39304; -. DR OMA; WEQAQEW; -. DR OrthoDB; 43475at2; -. DR PhylomeDB; P39304; -. DR BioCyc; EcoCyc:G7858-MONOMER; -. DR BioCyc; MetaCyc:G7858-MONOMER; -. DR BRENDA; 4.1.1.85; 2026. DR SABIO-RK; P39304; -. DR UniPathway; UPA00263; UER00378. DR EvolutionaryTrace; P39304; -. DR PRO; PR:P39304; -. DR Proteomes; UP000000318; Chromosome. DR Proteomes; UP000000625; Chromosome. DR GO; GO:0033982; F:3-dehydro-L-gulonate-6-phosphate decarboxylase activity; IDA:EcoCyc. DR GO; GO:0000287; F:magnesium ion binding; IDA:EcoCyc. DR GO; GO:0004590; F:orotidine-5'-phosphate decarboxylase activity; IEA:InterPro. DR GO; GO:0006207; P:'de novo' pyrimidine nucleobase biosynthetic process; IEA:InterPro. DR GO; GO:0019854; P:L-ascorbic acid catabolic process; IEP:EcoCyc. DR CDD; cd04726; KGPDC_HPS; 1. DR Gene3D; 3.20.20.70; Aldolase class I; 1. DR HAMAP; MF_01267; UlaD; 1. DR InterPro; IPR023942; 3-keto-L-gulonate6Pdecase_UlaD. DR InterPro; IPR013785; Aldolase_TIM. DR InterPro; IPR041710; HPS/KGPDC. DR InterPro; IPR001754; OMPdeCOase_dom. DR InterPro; IPR011060; RibuloseP-bd_barrel. DR PANTHER; PTHR35039; 3-KETO-L-GULONATE-6-PHOSPHATE DECARBOXYLASE SGBH-RELATED; 1. DR PANTHER; PTHR35039:SF3; 3-KETO-L-GULONATE-6-PHOSPHATE DECARBOXYLASE SGBH-RELATED; 1. DR Pfam; PF00215; OMPdecase; 1. DR SMART; SM00934; OMPdecase; 1. DR SUPFAM; SSF51366; Ribulose-phoshate binding barrel; 1. DR SWISS-2DPAGE; P39304; -. PE 1: Evidence at protein level; KW 3D-structure; Carbohydrate metabolism; Decarboxylase; Lyase; Magnesium; KW Metal-binding; Reference proteome. FT CHAIN 1..216 FT /note="3-keto-L-gulonate-6-phosphate decarboxylase UlaD" FT /id="PRO_0000212103" FT BINDING 11 FT /ligand="substrate" FT BINDING 33 FT /ligand="Mg(2+)" FT /ligand_id="ChEBI:CHEBI:18420" FT BINDING 62 FT /ligand="Mg(2+)" FT /ligand_id="ChEBI:CHEBI:18420" FT BINDING 192 FT /ligand="substrate" FT SITE 64 FT /note="Transition state stabilizer" FT SITE 67 FT /note="Transition state stabilizer" FT MUTAGEN 33 FT /note="E->K: Loss of activity." FT /evidence="ECO:0000269|PubMed:15157077" FT MUTAGEN 64 FT /note="K->A: 16% of wild-type activity." FT /evidence="ECO:0000269|PubMed:15157077" FT MUTAGEN 67 FT /note="D->A: 5% of wild-type activity." FT /evidence="ECO:0000269|PubMed:15157077" FT MUTAGEN 112 FT /note="E->A: 0.5% of wild-type activity." FT /evidence="ECO:0000269|PubMed:15157077" FT MUTAGEN 136 FT /note="H->A: 5% of wild-type activity." FT /evidence="ECO:0000269|PubMed:15157077" FT MUTAGEN 139 FT /note="R->V: 17% of wild-type activity." FT /evidence="ECO:0000269|PubMed:15157077" FT STRAND 5..10 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 15..25 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 26..28 FT /evidence="ECO:0007829|PDB:1Q6O" FT STRAND 30..34 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 36..42 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 45..53 FT /evidence="ECO:0007829|PDB:1Q6O" FT STRAND 57..65 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 69..78 FT /evidence="ECO:0007829|PDB:1Q6O" FT STRAND 82..87 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 92..104 FT /evidence="ECO:0007829|PDB:1Q6O" FT STRAND 108..113 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 119..127 FT /evidence="ECO:0007829|PDB:1Q6O" FT STRAND 132..136 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 139..143 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 150..161 FT /evidence="ECO:0007829|PDB:1Q6O" FT STRAND 165..171 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 174..180 FT /evidence="ECO:0007829|PDB:1Q6O" FT STRAND 186..191 FT /evidence="ECO:0007829|PDB:1Q6O" FT HELIX 192..195 FT /evidence="ECO:0007829|PDB:1Q6O" FT STRAND 197..199 FT /evidence="ECO:0007829|PDB:1KV8" FT HELIX 200..214 FT /evidence="ECO:0007829|PDB:1Q6O" SQ SEQUENCE 216 AA; 23578 MW; EC8490DA1D02D824 CRC64; MSLPMLQVAL DNQTMDSAYE TTRLIAEEVD IIEVGTILCV GEGVRAVRDL KALYPHKIVL ADAKIADAGK ILSRMCFEAN ADWVTVICCA DINTAKGALD VAKEFNGDVQ IELTGYWTWE QAQQWRDAGI GQVVYHRSRD AQAAGVAWGE ADITAIKRLS DMGFKVTVTG GLALEDLPLF KGIPIHVFIA GRSIRDAASP VEAARQFKRS IAELWG //