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P06632 (DKGA_CORSC) Reviewed, UniProtKB/Swiss-Prot

Last modified February 19, 2014. Version 83. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
2,5-diketo-D-gluconic acid reductase A

Short name=2,5-DKG reductase A
Short name=2,5-DKGR A
Short name=25DKGR-A
EC=1.1.1.346
Alternative name(s):
AKR5C
Gene names
Name:dkgA
OrganismCorynebacterium sp. (strain ATCC 31090)
Taxonomic identifier268952 [NCBI]
Taxonomic lineageBacteriaActinobacteriaActinobacteridaeActinomycetalesCorynebacterineaeCorynebacteriaceaeCorynebacterium

Protein attributes

Sequence length278 AA.
Sequence statusComplete.
Sequence processingThe displayed sequence is further processed into a mature form.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

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 activity

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

Enzyme regulation

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

Subunit structure

Monomer.

Subcellular location

Cytoplasm.

Biotechnological use

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.

Miscellaneous

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

Sequence similarities

Belongs to the aldo/keto reductase family.

Biophysicochemical properties

Kinetic parameters:

KM=26 mM for 2,5-diketo-D-gluconate Ref.2

KM=10 µM for NADPH

pH dependence:

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

Ontologies

Keywords
   Biological processAscorbate biosynthesis
   Cellular componentCytoplasm
   LigandNADP
   Molecular functionOxidoreductase
   Technical term3D-structure
Direct protein sequencing
Gene Ontology (GO)
   Biological_processL-ascorbic acid biosynthetic process

Inferred from electronic annotation. Source: UniProtKB-KW

   Cellular_componentcytoplasm

Inferred from electronic annotation. Source: UniProtKB-SubCell

   Molecular_functionoxidoreductase activity

Inferred from electronic annotation. Source: UniProtKB-KW

Complete GO annotation...

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Initiator methionine11Removed Ref.1 Ref.2
Chain2 – 2782772,5-diketo-D-gluconic acid reductase A
PRO_0000124597

Regions

Nucleotide binding188 – 24255NADP

Sites

Active site501Proton donor
Binding site1081Substrate

Experimental info

Mutagenesis221F → Y: Tighter binding of NADH.
Mutagenesis2321K → G: Tighter binding of NADH.
Mutagenesis2381R → H: Tighter binding of NADH.
Mutagenesis2721A → G: Tighter binding of NADH.

Secondary structure

.................................................... 278
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P06632 [UniParc].

Last modified January 23, 2007. Version 3.
Checksum: 403A2E9AA4624A37

FASTA27830,119
        10         20         30         40         50         60 
MTVPSIVLND GNSIPQLGYG VFKVPPADTQ RAVEEALEVG YRHIDTAAIY GNEEGVGAAI 

        70         80         90        100        110        120 
AASGIARDDL FITTKLWNDR HDGDEPAAAI AESLAKLALD QVDLYLVHWP TPAADNYVHA 

       130        140        150        160        170        180 
WEKMIELRAA GLTRSIGVSN HLVPHLERIV AATGVVPAVN QIELHPAYQQ REITDWAAAH 

       190        200        210        220        230        240 
DVKIESWGPL GQGKYDLFGA EPVTAAAAAH GKTPAQAVLR WHLQKGFVVF PKSVRRERLE 

       250        260        270 
ENLDVFDFDL TDTEIAAIDA MDPGDGSGRV SAHPDEVD 

« Hide

References

[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.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
M12799 Genomic DNA. Translation: AAA83534.1.
PIRI40838.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1A80X-ray2.10A2-278[»]
1HW6X-ray1.90A1-278[»]
1M9HX-ray2.00A1-278[»]
ProteinModelPortalP06632.
SMRP06632. Positions 2-278.
ModBaseSearch...
MobiDBSearch...

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Enzyme and pathway databases

BioCycMetaCyc:MONOMER-17833.
SABIO-RKP06632.

Family and domain databases

Gene3D3.20.20.100. 1 hit.
InterProIPR001395. Aldo/ket_red.
IPR018170. Aldo/ket_reductase_CS.
IPR020471. Aldo/keto_reductase_subgr.
IPR023210. NADP_OxRdtase_dom.
[Graphical view]
PANTHERPTHR11732. PTHR11732. 1 hit.
PfamPF00248. Aldo_ket_red. 1 hit.
[Graphical view]
PIRSFPIRSF000097. AKR. 1 hit.
PRINTSPR00069. ALDKETRDTASE.
SUPFAMSSF51430. SSF51430. 1 hit.
PROSITEPS00798. ALDOKETO_REDUCTASE_1. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceP06632.

Entry information

Entry nameDKGA_CORSC
AccessionPrimary (citable) accession number: P06632
Entry history
Integrated into UniProtKB/Swiss-Prot: November 1, 1988
Last sequence update: January 23, 2007
Last modified: February 19, 2014
This is version 83 of the entry and version 3 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references