DKGA_CORSC
ID DKGA_CORSC Reviewed; 278 AA.
AC P06632;
DT 01-NOV-1988, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 3.
DT 03-AUG-2022, entry version 111.
DE RecName: Full=2,5-diketo-D-gluconic acid reductase A;
DE Short=2,5-DKG reductase A;
DE Short=2,5-DKGR A;
DE Short=25DKGR-A;
DE EC=1.1.1.346;
DE AltName: Full=AKR5C;
GN Name=dkgA;
OS Corynebacterium sp. (strain ATCC 31090).
OC Bacteria; Actinobacteria; Corynebacteriales; Corynebacteriaceae;
OC Corynebacterium; unclassified Corynebacterium.
OX NCBI_TaxID=268952;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-41; 76-89; 97-123
RP AND 184-194, AND CHARACTERIZATION.
RA Anderson S., Marks C.B., Lazarus R.A., Miller J.V., Stafford K.,
RA Seymour J., Light D., Rastetter W., Estell D.A.;
RT "Production of 2-keto-L-gulonate, an intermediate in L-ascorbate synthesis,
RT by a genetically modified Erwinia herbicola.";
RL Science 230:144-149(1985).
RN [2]
RP PROTEIN SEQUENCE OF 2-41, CHARACTERIZATION, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RX PubMed=3597405; DOI=10.1016/s0021-9258(18)48039-8;
RA Miller J.V., Estell D.A., Lazarus R.A.;
RT "Purification and characterization of 2,5-diketo-D-gluconate reductase from
RT Corynebacterium sp.";
RL J. Biol. Chem. 262:9016-9020(1987).
RN [3]
RP 3D-STRUCTURE MODELING OF SUBSTRATE-BINDING SITE.
RX PubMed=10737928;
RX DOI=10.1002/(sici)1097-0134(20000401)39:1<68::aid-prot7>3.0.co;2-y;
RA Khurana S., Sanli G., Powers D.B., Anderson S., Blaber M.;
RT "Molecular modeling of substrate binding in wild-type and mutant
RT Corynebacteria 2,5-diketo-D-gluconate reductases.";
RL Proteins 39:68-75(2000).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS).
RX PubMed=9618487; DOI=10.1073/pnas.95.12.6768;
RA Khurana S., Powers D.B., Anderson S., Blaber M.;
RT "Crystal structure of 2,5-diketo-D-gluconic acid reductase A complexed with
RT NADPH at 2.1-A resolution.";
RL Proc. Natl. Acad. Sci. U.S.A. 95:6768-6773(1998).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS), AND ACTIVE SITE.
RX PubMed=11399090; DOI=10.1006/jmbi.2001.4739;
RA Sanli G., Blaber M.;
RT "Structural assembly of the active site in an aldo-keto reductase by NADPH
RT cofactor.";
RL J. Mol. Biol. 309:1209-1218(2001).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF MUTANT WITH TYR-22; GLY-232;
RP HIS-238 AND GLY-272 IN COMPLEX WITH NADH.
RX PubMed=14718658; DOI=10.1110/ps.03450704;
RA Sanli G., Banta S., Anderson S., Blaber M.;
RT "Structural alteration of cofactor specificity in Corynebacterium 2,5-
RT diketo-D-gluconic acid reductase.";
RL Protein Sci. 13:504-512(2004).
CC -!- FUNCTION: Catalyzes the reduction of 2,5-diketo-D-gluconic acid (25DKG)
CC to 2-keto-L-gulonic acid (2KLG). 5-keto-D-fructose and dihydroxyacetone
CC can also serve as substrates. 25DKGR-A exhibits a greater selectivity
CC for the substrate and higher thermal stability than 25DKGR-B.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-dehydro-L-idonate + NADP(+) = 2,5-didehydro-D-gluconate +
CC H(+) + NADPH; Xref=Rhea:RHEA:35111, ChEBI:CHEBI:11449,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:36602, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; EC=1.1.1.346;
CC -!- ACTIVITY REGULATION: Inhibited by Zn(2+), Fe(3+), Cu(2+) and Ni(2+).
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=26 mM for 2,5-diketo-D-gluconate {ECO:0000269|PubMed:3597405};
CC KM=10 uM for NADPH {ECO:0000269|PubMed:3597405};
CC pH dependence:
CC Optimum pH is 6.4. Active over a broad pH range.
CC {ECO:0000269|PubMed:3597405};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:14718658}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- BIOTECHNOLOGY: Introduced by genetic manipulation and expressed in
CC Erwinia herbicola by Anderson et al. The resultant organism is able to
CC convert in a single fermentative step D-glucose into 2-keto-L-gulonic
CC acid, a key precursor in the industrial production of L-ascorbic acid
CC (vitamin C). However, the technology still needs some working on and is
CC not used in commercial production at present.
CC -!- MISCELLANEOUS: Modeling study indicates that the active site may not be
CC optimized for 2,5-diketo-D-gluconic acid.
CC -!- SIMILARITY: Belongs to the aldo/keto reductase family. {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; M12799; AAA83534.1; -; Genomic_DNA.
DR PIR; I40838; I40838.
DR PDB; 1A80; X-ray; 2.10 A; A=2-278.
DR PDB; 1HW6; X-ray; 1.90 A; A=1-278.
DR PDB; 1M9H; X-ray; 2.00 A; A=1-278.
DR PDBsum; 1A80; -.
DR PDBsum; 1HW6; -.
DR PDBsum; 1M9H; -.
DR AlphaFoldDB; P06632; -.
DR SMR; P06632; -.
DR DrugBank; DB03461; Nicotinamide adenine dinucleotide phosphate.
DR KEGG; ag:AAA83534; -.
DR BioCyc; MetaCyc:MON-17833; -.
DR BRENDA; 1.1.1.274; 1664.
DR BRENDA; 1.1.1.346; 1664.
DR SABIO-RK; P06632; -.
DR EvolutionaryTrace; P06632; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR GO; GO:0019853; P:L-ascorbic acid biosynthetic process; IEA:UniProtKB-KW.
DR CDD; cd19130; AKR_AKR5C1; 1.
DR Gene3D; 3.20.20.100; -; 1.
DR InterPro; IPR020471; AKR.
DR InterPro; IPR044503; AKR5C1.
DR InterPro; IPR018170; Aldo/ket_reductase_CS.
DR InterPro; IPR023210; NADP_OxRdtase_dom.
DR InterPro; IPR036812; NADP_OxRdtase_dom_sf.
DR PANTHER; PTHR43827; PTHR43827; 1.
DR Pfam; PF00248; Aldo_ket_red; 1.
DR PIRSF; PIRSF000097; AKR; 1.
DR PRINTS; PR00069; ALDKETRDTASE.
DR SUPFAM; SSF51430; SSF51430; 1.
DR PROSITE; PS00798; ALDOKETO_REDUCTASE_1; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Ascorbate biosynthesis; Cytoplasm; Direct protein sequencing;
KW NADP; Oxidoreductase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:3597405, ECO:0000269|Ref.1"
FT CHAIN 2..278
FT /note="2,5-diketo-D-gluconic acid reductase A"
FT /id="PRO_0000124597"
FT REGION 259..278
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 50
FT /note="Proton donor"
FT /evidence="ECO:0000269|PubMed:11399090"
FT BINDING 108
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:11399090"
FT BINDING 188..242
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:14718658"
FT MUTAGEN 22
FT /note="F->Y: Tighter binding of NADH."
FT MUTAGEN 232
FT /note="K->G: Tighter binding of NADH."
FT MUTAGEN 238
FT /note="R->H: Tighter binding of NADH."
FT MUTAGEN 272
FT /note="A->G: Tighter binding of NADH."
FT STRAND 5..7
FT /evidence="ECO:0007829|PDB:1HW6"
FT STRAND 13..17
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 26..28
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 29..39
FT /evidence="ECO:0007829|PDB:1HW6"
FT STRAND 43..45
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 47..49
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 54..63
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 67..69
FT /evidence="ECO:0007829|PDB:1HW6"
FT STRAND 71..76
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 78..80
FT /evidence="ECO:0007829|PDB:1M9H"
FT HELIX 86..97
FT /evidence="ECO:0007829|PDB:1HW6"
FT STRAND 102..107
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 112..114
FT /evidence="ECO:0007829|PDB:1M9H"
FT HELIX 117..129
FT /evidence="ECO:0007829|PDB:1HW6"
FT STRAND 132..140
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 143..153
FT /evidence="ECO:0007829|PDB:1HW6"
FT STRAND 158..163
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 171..179
FT /evidence="ECO:0007829|PDB:1HW6"
FT STRAND 183..188
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 191..193
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 201..210
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 214..224
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 236..243
FT /evidence="ECO:0007829|PDB:1HW6"
FT HELIX 252..259
FT /evidence="ECO:0007829|PDB:1HW6"
FT STRAND 264..266
FT /evidence="ECO:0007829|PDB:1M9H"
FT TURN 274..276
FT /evidence="ECO:0007829|PDB:1M9H"
SQ SEQUENCE 278 AA; 30119 MW; 403A2E9AA4624A37 CRC64;
MTVPSIVLND GNSIPQLGYG VFKVPPADTQ RAVEEALEVG YRHIDTAAIY GNEEGVGAAI
AASGIARDDL FITTKLWNDR HDGDEPAAAI AESLAKLALD QVDLYLVHWP TPAADNYVHA
WEKMIELRAA GLTRSIGVSN HLVPHLERIV AATGVVPAVN QIELHPAYQQ REITDWAAAH
DVKIESWGPL GQGKYDLFGA EPVTAAAAAH GKTPAQAVLR WHLQKGFVVF PKSVRRERLE
ENLDVFDFDL TDTEIAAIDA MDPGDGSGRV SAHPDEVD
