Information on EC 5.1.3.1 - ribulose-phosphate 3-epimerase

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Substrates: reaction of EC 5.1.3.1
Products: -

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D-Ribulose 5-phosphate

D-Xylulose 5-phosphate

32 entries

D-ribulose-5-phosphate

D-xylulose 5-phosphate

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Substrates: -
Products: -

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D-xylulose 5-phosphate

D-ribulose 5-phosphate

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662486

Substrates: -
Products: -

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additional information

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6 entries

D-Ribulose 5-phosphate

D-Xylulose 5-phosphate

14 entries

additional information

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-

6 entries

Co2+

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Co2+ is able to activate the enzyme in vitro

716784

Fe2+

Mg2+

assay carried out at 10 mM Mg2+

Mn2+

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manganese can activate the enzyme in vitro in place of iron converting the enzyme to a form that is unaffected by H2O2

716784

Zn2+

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Zn2+ is able to activate the enzyme in vitro

716784

additional information

3 entries

Co2+

I3DTN4, I3DZ65, I3DTP3

causes over 80% inhibition at 5 mM; causes over 80% inhibition at 5 mM; causes over 80% inhibition at 5 mM

Cu(CH3COO)2

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D-deoxyribose 5-phosphate

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DL-alpha-glycerophosphate

competitive

HgCl2

iodoacetate

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weak

Mg2+

I3DTN4, I3DZ65, I3DTP3

causes only slight inhibition at 5 mM; causes only slight inhibition at 5 mM; causes only slight inhibition at 5 mM

Mn2+

I3DTN4, I3DZ65, I3DTP3

causes 20% inhibition at 5 mM; causes 20% inhibition at 5 mM; causes 20% inhibition at 5 mM

NEM

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weak

PCMB

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potassium phosphate

SO42-

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Sodium sulfate

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20 mM, 65% inhibition of D-ribulose-5-phosphate 3-epimerase activity

Zn2+

I3DTN4, I3DZ65, I3DTP3

causes complete inhibition at 5 mM; causes complete inhibition at 5 mM; causes complete inhibition at 5 mM

additional information

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Zn2+

RPE is activated by Zn2+ which binds with a stoichiometry of one ion per polypeptide

0.056 - 15

D-ribulose 5-phosphate

14 entries

2.4 - 2.9

D-ribulose-5-phosphate

additional information

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0.00033 - 7100

D-ribulose 5-phosphate

12 entries

1300

D-xylulose 5-phosphate

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2225

0.9

DL-alpha-glycerophosphate

25°C, pH 8.0

11610

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6570

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substrate D-ribulose-5-phosphate, pH 7.0, 30°C

9.05

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substrate D-erythrose 4-phosphate, pH 7.0, 30°C

additional information

7.4 - 7.6

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7.5

assay at

7.7

I3DTN4, I3DZ65, I3DTP3

assay at

8

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8.3

8.5

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6.5 - 9

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pH 6.5: about 90% of maximal activity, pH 9.0: 95% of maximal activity

2239

25

I3DTN4, I3DZ65, I3DTP3

assay at

4.8

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isoelectric focusing

6.3 - 6.8

wild-type enzyme and mutant enzyme D186E, denaturing isoelectric focusing, pH gradient 5-8

6.6

mutant enzymes D186A and D186N, denaturing isoelectric focusing, pH gradient 5-8

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SwissProt

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I3DTN4, I3DTP3, I3DZ65

UniProt

Bacillus methanolicus MGA3 / ATCC53907

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I3DTN4, I3DZ65

UniProt

Bacillus methanolicus PB1 / NCIMB13113

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Bacillus sp. (in: firmicutes)

I3DTP3

UniProt

mutant strain that lacks D-ribulose-5-phosphate 3-epimerase

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SwissProt

Bacillus subtilis 168

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SwissProt

-

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basonym Alcaligenes eutrophus

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-

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Lacticaseibacillus casei

Lacticaseibacillus casei 64H

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727770

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Lathyrus oleraceus

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676674

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Oryctolagus cuniculus

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2228

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Pigeon

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2228

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Rattus norvegicus

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2228, 2233

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Saccharomyces cerevisiae ATCC 204508 / S288c

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UniProt

-

-

Synechocystis sp. PCC 6803

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Uniprot

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UniProt

-

-

-

-

additional information

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nearest neighbor analyses of immunocytolocalization experiments indicate that glyceraldehyde-3-P dehydrogenase and phosphoribulokinase are located close to xylulose-5-P 3-epimerase in the pea leaf chloroplast stroma

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Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

evolution

malfunction

Chlamydomonas reinhardtii

metabolism

7 entries

physiological function

Francisella tularensis subsp. tularensis

Histophilus somni 129PT

Neisseria gonorrhoeae NCCP11945

Pseudomonas aeruginosa PAO1

Stenotrophomonas maltophilia K279a

180000

-

gel filtration

200000

gel filtration

22500

-

2 * 22500, SDS-PAGE

22900

-

2 * 22900, SDS-PAGE

23000

24300

2 * 24300, cyt-RPEase, calculation from nucleotide sequence

25000

8 * 25000, SDS-PAGE

25966

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x * 25966, calculation from nucleotide sequence

2241

26000

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2 * 26000, SDS-PAGE

2236

43000

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gel filtration

45000

45900

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ultracentrifugation

2229

50000

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sedimentation equilibrium experiments

2236

55000

cyt-RPEase, gel filtration

651988

?

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x * 25966, calculation from nucleotide sequence

dimer

hexamer

octamer

hanging drop vapor diffusion method, using 25% (w/v) PEG 3350, 100 mM Bis-Tris (pH 5.5), and 200 mM NaCl

crystals are produced with hanging drop method using 6 mg/ml cytosolic enzyme, 1.0 M ammonium sulfate, 50 mM Mops, pH 7.5, 1% w/v polyethylene glycol 400, 10 mM glycerol-3-phosphate and 5 mM L-arginine in the drop

652903

hanging drop vapor diffusion method

Solanum tuberosum

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652793

crystal structure of the RPE is solved at 1.8 A resolution in the presence of D-xylitol 5-phosphate, an inert analogue of the D-xylulose 5-phosphate substrate. This structure suggests that the 2,3-enediolate intermediate in the 1,1-proton transfer reaction is stabilized by bidentate coordination to the Zn2+ that also is liganded to His 34, Asp 36, His 67, and Asp 176, the carboxylate groups of the Asp residues are positioned also to function as the acid/base catalysts

crystals are grown using the microbatch technique from 22% methyl ether PEG 2000, 200 mM MgCl2, 100 mM MES pH 6.0. The crystals belong tp space group P2(1), with unit-cell parameters a = 82.6 A, b = 87.8 A, c = 97.8 A, beta = 114.6°, 1.6 A resolution

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D175A

inactive

D37A

the mutation almost abolishes the enzymatic activity

H35A

the mutation almost abolishes the enzymatic activity

H70A

completely insoluble

L12A

the mutation results in more than 50% decrease in the activity

M141A

the mutation results in about 30% decrease in the activity

M39A

the mutation results in about 10% decrease in the activity

M72A

the mutation results in an almost 50% decrease in the activity

S10A

the mutation almost abolishes the enzymatic activity

D186A

KM-value for D-ribulose 5-phosphate is increased 10fold, turnover-number is decreased to less than 0.1% of the wild-type value

D186E

KM-value for D-ribulose 5-phosphate is unaltered, turnover-number is decreased to less than 0.1% of the wild-type value

D186N

KM-value for D-ribulose 5-phosphate is unaltered, turnover-number is decreased to less than 0.1% of the wild-type value

D176A

no detectable activity, substrate= 10 mM D-ribulose 5-phosphate

D36A

no detectable activity, substrate= 10 mM D-ribulose 5-phosphate

H34A

mutant discloses decreased affinity for Zn2+, kcat = 200/sec, substrate = 10 mM D-ribulose 5-phosphate, +10 micromol/l ZnCl2

H67A

mutant discloses decreased affinity for Zn2+, kcat = 270/sec, substrate = 10 mM D-ribulose 5-phosphate, +10 micromol/l ZnCl2

additional information

6 entries

5.5

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24 h, 25°C, about 70% loss of activity

6.5 - 9.5

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22 h, 0°C, stable

7 - 8

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24 h, 25°C, stable

9

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24 h, 25°C, about 20% loss of activity

27

I3DTN4, I3DZ65, I3DTP3

purified recombinant His-tagged RPE2-MGA3 is completely stable at 27°C after 2 h

27 - 80

37

I3DTN4, I3DZ65, I3DTP3

purified recombinant His-tagged RPE2-MGA3 retains 95% activity after 2 h

50

I3DTN4, I3DZ65, I3DTP3

purified recombinant His-tagged RPE2-MGA3 retains 90% activity after 2 h

60

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2 min, stable

65

80

I3DTN4, I3DZ65, I3DTP3

purified recombinant His-tagged RPE2-MGA3 retains below 5% activity after 2 h

additional information

DL-alpha-glycerophosphate or ethanol stabilize the extremely labile recombinant enzyme, but are unable to reverse the spontaneous loss of activity

653499

drastically destabilized by 2-mercaptoethanol

DTT destabilizes

ribulose-5-phosphate 3-epimerase is vulnerable to H2O2 damage in vivo. H2O2 rapidly oxidizes the Fe2+ in a Fenton reaction, the oxidized iron is released immediately, causing a loss of activity. When purified enzyme is loaded with Mn2+, Co2+, or Zn2+, there is no loss in activity when challenged with H2O2

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716784

-20°C, 0.5 M ammonium sulfate, stable for at least 2 months

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2229

2°C, wether in crude extracts or highly purified, the recombinant enzyme is inherently unstable

653499

4°C, 10 mM Bistris/HCl buffer (pH 7.0), stable for about six months without any loss of enzymatic activity

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4°C, stable as ammonium precipitate for several weeks

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2236

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lysate is applied to a chelating sepharose fast flow column charged with Ni2+, the N-terminal His tag is removed by thrombin cleavage

mutant enzymes D186A, D186N, D186E

Ni-affinity column chromatography and Superdex G75 gel filtration

partial

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recombinant cytosolic enzyme

651988

recombinant His-tagged enzyme

Lacticaseibacillus casei

A0A510WIR9

727770

recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and diaylsis

I3DTN4, I3DZ65, I3DTP3

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DNA and amino acid sequence determination and analysis, expression as His-tagged enzyme

Lacticaseibacillus casei

A0A510WIR9

727770

enzyme with half normal activity in a malformed and severely mentally retarded girl with a de novo interstitial deletion 46,XX.del(2)(q32.lq34). It is suggested that the gene for ribulose-phosphate 3-epimerase is located on the segment 2q32.1 to q34

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2234, 2235

Escherichia coli clones harboring the cfxE gene show up to 19-fold-higher activities of ribulose-phosphate 3-epimerase

Cupriavidus necator

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2244

expressed in Escherichia coli BL21(DE3) cells

expression in Escherichia coli

expression of cytosoli enzyme in Escherichia coli

Synechocystis sp. PCC 6803

651988

gene rpe, recombinant overexpression in Synechocystis sp. PCC 6803

P74061

749337

gene rpe1 or rpeP is plasmid-encoded in Bacillus methanolicus strain MGA3, DNA and amino acid sequence determination and analysis, genomic organization, sequence comparisons and phylogenetic tree, real-time PCR enzyme expression analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)

I3DTN4, I3DZ65, I3DTP3

gene rpe1 or rpeP is plasmid-encoded in Bacillus methanolicus strain PB1, DNA and amino acid sequence determination and analysis, genomic organization, sequence comparisons and phylogenetic tree, real-time PCR enzyme expression analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)

I3DTN4, I3DZ65, I3DTP3

gene RPE1, subcloning in Escherichia coli DH5alpha, recombinant expression in Saccharomyces cerevisiae strain L2612

P46969

747383

gene rpe2 or rpeC is encoded on the chromosome in Bacillus methanolicus strain MGA3, DNA and amino acid sequence determination and analysis, genomic organization, sequence comparisons and phylogenetic tree, real-time PCR enzyme expression analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)

I3DTN4, I3DZ65, I3DTP3

the enzyme for ribulose 5-phosphate epimerase is localized to chromosome X

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2241

the gene encoding RPE is expressed in the Escherichia coli strain BL21 (DE3)