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Protein

2,5-diketo-D-gluconic acid reductase A

Gene

dkgA

Organism
Corynebacterium sp. (strain ATCC 31090)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the reduction of 2,5-diketo-D-gluconic acid (25DKG) to 2-keto-L-gulonic acid (2KLG). 5-keto-D-fructose and dihydroxyacetone can also serve as substrates. 25DKGR-A exhibits a greater selectivity for the substrate and higher thermal stability than 25DKGR-B.

Catalytic activityi

2-dehydro-L-gulonate + NADP+ = 2,5-didehydro-D-gluconate + NADPH.

Enzyme regulationi

Inhibited by Zn2+, Fe3+, Cu2+ and Ni2+.

Kineticsi

  1. KM=26 mM for 2,5-diketo-D-gluconate1 Publication
  2. KM=10 µM for NADPH1 Publication

pH dependencei

Optimum pH is 6.4. Active over a broad pH range.1 Publication

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Active sitei50 – 501Proton donor
Binding sitei108 – 1081Substrate

Regions

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Nucleotide bindingi188 – 24255NADP1 PublicationAdd
BLAST

GO - Molecular functioni

  1. oxidoreductase activity Source: UniProtKB-KW

GO - Biological processi

  1. L-ascorbic acid biosynthetic process Source: UniProtKB-KW
Complete GO annotation...

Keywords - Molecular functioni

Oxidoreductase

Keywords - Biological processi

Ascorbate biosynthesis

Keywords - Ligandi

NADP

Enzyme and pathway databases

BioCyciMetaCyc:MONOMER-17833.
SABIO-RKP06632.

Names & Taxonomyi

Protein namesi
Recommended name:
2,5-diketo-D-gluconic acid reductase A (EC:1.1.1.346)
Short name:
2,5-DKG reductase A
Short name:
2,5-DKGR A
Short name:
25DKGR-A
Alternative name(s):
AKR5C
Gene namesi
Name:dkgA
OrganismiCorynebacterium sp. (strain ATCC 31090)
Taxonomic identifieri268952 [NCBI]
Taxonomic lineageiBacteriaActinobacteriaActinobacteridaeActinomycetalesCorynebacterineaeCorynebacteriaceaeCorynebacterium

Subcellular locationi

GO - Cellular componenti

  1. cytoplasm Source: UniProtKB-SubCell
Complete GO annotation...

Keywords - Cellular componenti

Cytoplasm

Pathology & Biotechi

Biotechnological usei

Introduced by genetic manipulation and expressed in Erwinia herbicola by Anderson et al. The resultant organism is able to convert in a single fermentative step D-glucose into 2-keto-L-gulonic acid, a key precursor in the industrial production of L-ascorbic acid (vitamin C). However, the technology still needs some working on and is not used in commercial production at present.

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi22 – 221F → Y: Tighter binding of NADH.
Mutagenesisi232 – 2321K → G: Tighter binding of NADH.
Mutagenesisi238 – 2381R → H: Tighter binding of NADH.
Mutagenesisi272 – 2721A → G: Tighter binding of NADH.

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Initiator methioninei1 – 11Removed2 Publications
Chaini2 – 2782772,5-diketo-D-gluconic acid reductase APRO_0000124597Add
BLAST

Interactioni

Subunit structurei

Monomer.1 Publication

Structurei

Secondary structure

1
278
Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Beta strandi5 – 73Combined sources
Beta strandi13 – 175Combined sources
Helixi26 – 283Combined sources
Helixi29 – 3911Combined sources
Beta strandi43 – 453Combined sources
Helixi47 – 493Combined sources
Helixi54 – 6310Combined sources
Helixi67 – 693Combined sources
Beta strandi71 – 766Combined sources
Helixi78 – 803Combined sources
Helixi86 – 9712Combined sources
Beta strandi102 – 1076Combined sources
Helixi112 – 1143Combined sources
Helixi117 – 12913Combined sources
Beta strandi132 – 1409Combined sources
Helixi143 – 15311Combined sources
Beta strandi158 – 1636Combined sources
Helixi171 – 1799Combined sources
Beta strandi183 – 1886Combined sources
Helixi191 – 1933Combined sources
Helixi201 – 21010Combined sources
Helixi214 – 22411Combined sources
Helixi236 – 2438Combined sources
Helixi252 – 2598Combined sources
Beta strandi264 – 2663Combined sources
Turni274 – 2763Combined sources

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1A80X-ray2.10A2-278[»]
1HW6X-ray1.90A1-278[»]
1M9HX-ray2.00A1-278[»]
ProteinModelPortaliP06632.
SMRiP06632. Positions 2-278.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiP06632.

Family & Domainsi

Sequence similaritiesi

Belongs to the aldo/keto reductase family.Curated

Family and domain databases

Gene3Di3.20.20.100. 1 hit.
InterProiIPR001395. Aldo/ket_red.
IPR018170. Aldo/ket_reductase_CS.
IPR020471. Aldo/keto_reductase_subgr.
IPR023210. NADP_OxRdtase_dom.
[Graphical view]
PANTHERiPTHR11732. PTHR11732. 1 hit.
PfamiPF00248. Aldo_ket_red. 1 hit.
[Graphical view]
PIRSFiPIRSF000097. AKR. 1 hit.
PRINTSiPR00069. ALDKETRDTASE.
SUPFAMiSSF51430. SSF51430. 1 hit.
PROSITEiPS00798. ALDOKETO_REDUCTASE_1. 1 hit.
[Graphical view]

Sequencei

Sequence statusi: Complete.

Sequence processingi: The displayed sequence is further processed into a mature form.

P06632-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MTVPSIVLND GNSIPQLGYG VFKVPPADTQ RAVEEALEVG YRHIDTAAIY
60 70 80 90 100
GNEEGVGAAI AASGIARDDL FITTKLWNDR HDGDEPAAAI AESLAKLALD
110 120 130 140 150
QVDLYLVHWP TPAADNYVHA WEKMIELRAA GLTRSIGVSN HLVPHLERIV
160 170 180 190 200
AATGVVPAVN QIELHPAYQQ REITDWAAAH DVKIESWGPL GQGKYDLFGA
210 220 230 240 250
EPVTAAAAAH GKTPAQAVLR WHLQKGFVVF PKSVRRERLE ENLDVFDFDL
260 270
TDTEIAAIDA MDPGDGSGRV SAHPDEVD
Length:278
Mass (Da):30,119
Last modified:January 23, 2007 - v3
Checksum:i403A2E9AA4624A37
GO

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
M12799 Genomic DNA. Translation: AAA83534.1.
PIRiI40838.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
M12799 Genomic DNA. Translation: AAA83534.1.
PIRiI40838.

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
1A80X-ray2.10A2-278[»]
1HW6X-ray1.90A1-278[»]
1M9HX-ray2.00A1-278[»]
ProteinModelPortaliP06632.
SMRiP06632. Positions 2-278.
ModBaseiSearch...
MobiDBiSearch...

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Enzyme and pathway databases

BioCyciMetaCyc:MONOMER-17833.
SABIO-RKP06632.

Miscellaneous databases

EvolutionaryTraceiP06632.

Family and domain databases

Gene3Di3.20.20.100. 1 hit.
InterProiIPR001395. Aldo/ket_red.
IPR018170. Aldo/ket_reductase_CS.
IPR020471. Aldo/keto_reductase_subgr.
IPR023210. NADP_OxRdtase_dom.
[Graphical view]
PANTHERiPTHR11732. PTHR11732. 1 hit.
PfamiPF00248. Aldo_ket_red. 1 hit.
[Graphical view]
PIRSFiPIRSF000097. AKR. 1 hit.
PRINTSiPR00069. ALDKETRDTASE.
SUPFAMiSSF51430. SSF51430. 1 hit.
PROSITEiPS00798. ALDOKETO_REDUCTASE_1. 1 hit.
[Graphical view]
ProtoNetiSearch...

Publicationsi

  1. "Production of 2-keto-L-gulonate, an intermediate in L-ascorbate synthesis, by a genetically modified Erwinia herbicola."
    Anderson S., Marks C.B., Lazarus R.A., Miller J.V., Stafford K., Seymour J., Light D., Rastetter W., Estell D.A.
    Science 230:144-149(1985)
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-41; 76-89; 97-123 AND 184-194, CHARACTERIZATION.
  2. "Purification and characterization of 2,5-diketo-D-gluconate reductase from Corynebacterium sp."
    Miller J.V., Estell D.A., Lazarus R.A.
    J. Biol. Chem. 262:9016-9020(1987) [PubMed] [Europe PMC] [Abstract]
    Cited for: PROTEIN SEQUENCE OF 2-41, CHARACTERIZATION, BIOPHYSICOCHEMICAL PROPERTIES.
  3. "Molecular modeling of substrate binding in wild-type and mutant Corynebacteria 2,5-diketo-D-gluconate reductases."
    Khurana S., Sanli G., Powers D.B., Anderson S., Blaber M.
    Proteins 39:68-75(2000) [PubMed] [Europe PMC] [Abstract]
    Cited for: 3D-STRUCTURE MODELING OF SUBSTRATE-BINDING SITE.
  4. "Crystal structure of 2,5-diketo-D-gluconic acid reductase A complexed with NADPH at 2.1-A resolution."
    Khurana S., Powers D.B., Anderson S., Blaber M.
    Proc. Natl. Acad. Sci. U.S.A. 95:6768-6773(1998) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS).
  5. "Structural assembly of the active site in an aldo-keto reductase by NADPH cofactor."
    Sanli G., Blaber M.
    J. Mol. Biol. 309:1209-1218(2001) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS).
  6. "Structural alteration of cofactor specificity in Corynebacterium 2,5-diketo-D-gluconic acid reductase."
    Sanli G., Banta S., Anderson S., Blaber M.
    Protein Sci. 13:504-512(2004) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF MUTANT WITH TYR-22; GLY-232; HIS-238 AND GLY-272 IN COMPLEX WITH NADH.

Entry informationi

Entry nameiDKGA_CORSC
AccessioniPrimary (citable) accession number: P06632
Entry historyi
Integrated into UniProtKB/Swiss-Prot: November 1, 1988
Last sequence update: January 23, 2007
Last modified: January 7, 2015
This is version 87 of the entry and version 3 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Miscellaneous

Modeling study indicates that the active site may not be optimized for 2,5-diketo-D-gluconic acid.

Keywords - Technical termi

3D-structure, Direct protein sequencing

Documents

  1. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references
  2. SIMILARITY comments
    Index of protein domains and families

External Data

Dasty 3

Similar proteinsi

Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into Uniref entry.
90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.