Information on EC 1.1.1.202 - 1,3-propanediol dehydrogenase

poor effects by EDTA, 2-mercaptoethanol, and sodium carbazide

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11.5

1,2-propanediol

pH 9.0, 60°C

26.7

1,3-Propanediol

pH 9.0, 60°C

0.77 - 10.05

3-hydroxypropanal

4.25

acetaldehyde

pH 7.0, 37°C

4.2

Butyraldehyde

pH 7.0, 37°C

16.1

formaldehyde

pH 7.0, 37°C

26 - 29

glyceraldehyde

300 - 700

glycerol

0.05 - 0.48

NAD+

11 entries

0.0116 - 0.48

NADH

9 entries

0.97

NADP+

mutant D41G, pH 7.4, 37°C

0.14

NADPH

mutant D41G, pH 7.4, 37°C

1.25 - 18

Propane-1,3-diol

9 entries

0.17 - 11

propionaldehyde

2.16

Valeraldehyde

pH 7.0, 37°C

additional information

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59.4

3-hydroxypropanal

pH 7.0, 37°C

5.2

acetaldehyde

pH 7.0, 37°C

8.7

Butyraldehyde

pH 7.0, 37°C

1.6

formaldehyde

pH 7.0, 37°C

9.7 - 82.33

NAD+

59.5 - 411.6

NADH

11.83

NADP+

mutant D41G, pH 7.4, 37°C

187.4

NADPH

mutant D41G, pH 7.4, 37°C

9.98

Propane-1,3-diol

pH 9.0, 55°C

1.1

propionaldehyde

pH 7.0, 37°C

Valeraldehyde

pH 7.0, 37°C

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0.01

1,2-propanediol

pH 9.0, 60°C

0.01

1,3-Propanediol

pH 9.0, 60°C

76.6

3-hydroxypropanal

pH 7.0, 37°C

1.2

acetaldehyde

pH 7.0, 37°C

2.1

Butyraldehyde

pH 7.0, 37°C

0.1

formaldehyde

pH 7.0, 37°C

4.9 - 260

NAD+

12.9 - 6043

NADH

12.2

NADP+

mutant D41G, pH 7.4, 37°C

1338

NADPH

mutant D41G, pH 7.4, 37°C

1.1

Propane-1,3-diol

pH 9.0, 55°C

1.1

propionaldehyde

pH 7.0, 37°C

0.5

Valeraldehyde

pH 7.0, 37°C

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12.09

glycerol

pH 9.0, 25°C

657406

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0.37

wild-type Clostridium butyricum strain VPI 3266

1.1

pH 9.0, 60°C

1.2

recombinant enzyme in Clostridium acetobutylicum mutant strain DG1

667270

1.7

Clostridium pasteurianum

389495

1.9

crude cell extract with recombinant His-tagged enzyme, substrate propane-1,3-diol, pH 9.0, 55°C

3.42

Pantoea agglomerans

pH and temperature not specified in the publication

3.8

native enzyme

675901

4.51

7.28

pH and temperature not specified in the publication

7.3

389490

9.6

recombinant enzyme

675901

9.85

pH and temperature not specified in the publication

pH and temperature not specified in the publication

13.1

purified recombinant His-tagged enzyme, substrate propane-1,3-diol, pH 9.0, 55°C

13.7

crude cell extract with recombinant His-tagged enzyme, substrate 3-hydroxypropanal, pH 7.0, 37°C

32.6

purified recombinant His-tagged enzyme, substrate propane-1,3-diol, pH 9.5, 25°C

49.5

reduction of 3-hydroxypropanal, pH 8.0, 37°C

52.7

purified recombinant His-tagged enzyme, substrate 3-hydroxypropanal, pH 7.5, 37°C

77.9

oxidation of propane-1,3-diol, pH 10.5, 25°C

89.7

purified recombinant His-tagged enzyme, substrate 3-hydroxypropanal, pH 7.0, 37°C

90.9

389494

additional information

nucleotide pools of wild-type Clostridium butyricum strain VPI 3266 and recombinant Clostridium acetobutylicum mutant strain DG1

667270

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6.2

3-hydroxypropanal + NADH

6.6

7.5

aldehyde reduction

7.7

7.8

9.07

3-hydroxypropanal + NADH

389496

9.1

alcohol oxidation

9.5

9.7

assay at

664422

9.72

propane-1,3-diol + NADH

389496

alcohol oxidation

10.5

oxidation of alcohol

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5 - 9.5

activity range, aldehyde reduction, profile overview

6 - 10

activity range, aldehyde reduction, profile overview

7 - 8.5

more than 80% of maximum activity for reduction of aldehyde

less than 10% of maximum activity for oxidation reaction

7 - 9

699357

8 - 10

activity range, alcohol oxidation, profile overview

8 - 10.5

activity range, alcohol oxidation, profile overview

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50 - 60

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15 - 40

activity range, aldehyde reduction, profile overview

25 - 45

activity range, alcohol oxidation, profile overview

30 - 55

over 80% of maximal activity within this range, aldehyde reduction, profile overview

30 - 65

over 60% of maximal activity within this range, alcohol oxidation, profile overview

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5.2

calculated

5.9

sequence calculation

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3bfj, download, 3D-view

SCOPe (3bfj)

CATH (3bfj)

Q7WRJ3

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38500

Pantoea agglomerans

41500

41544

x * 41544, sequence calculation

42000

42200

2 * 42200, calculated, 2 * 43000, SDS-PAGE of recombinant His-tagged protein

42500

43000

43400

45000

6 * or 8 * 45000, SDS-PAGE

46000

74000

gel filtration

180000

gel filtration

389490

328000

gel filtration

389488

331000

gel filtration, sedimentation coefficient

389494

336000

347000

350000

355000

384200

387000

415000

sequence analysis

440000

446000

dynamic light scattering analysis of the purified protein in solution

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5 entries

decamer

crystallography or gel filtration

dimer

hexamer or octamer

6 * or 8 * 45000, SDS-PAGE

389488

homodecamer

10 * 42927, sequence calculation, 10 * 43000, recombinant His6-tagged enzyme, SDS-PAGE

740927

monomer

Aquifex aeolicus

1 * 43000, calculated

710747

octamer

oligomer

8 entries

tetramer

4 * 42000, SDS-PAGE

389490

additional information

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selenomethionine-labeled protein, to 2.4 A resolution. space group P32

Aquifex aeolicus

710747

by vapor diffusion in sitting drops at 20°C, at 2.7 A resolution. The enzyme shows a decameric structure, formed by a pentamer of dimers, which is the catalytic form of the enzyme. Dimers are associated by strong ionic interactions that are responsible for the highly stable in vivo packing of the enzyme. The decameric arrangement is related to the cooperativity between monomers

homology modeling with cofactors NADH and NADPH

purified recombinant His-tagged enzyme, sitting-drop vapour diffusion, 22°C, 0.0015 ml of 55 mg/ml protein in 50 mM HEPES, pH 7.4, 150 mM NaCl, 1 mM MnCl2 and 2 mM DTT, is mixed with an equal volume of reservoir solution containing 0.1 M MES, pH 6.5, with 12% w/v PEG 20000, and 10 mM CaCl2, 8-24 h, X-ray diffraction structure determination and analysis at 2.7 A resolution

to 3.2 A resolution. The electron density map around the active site shows no sign of a bound co-factor, but the active site metal could be located and identified as Ni2+

purified recombinant His-tagged protein in 20 mM Tris, pH 7.9, 150 mM NaCl, 0.25 mM TCEP, by nandroplet vapour diffusion method, cyrstallization buffer contains 20% PEG 300, 5% w/v PEG 8000, 10% glycerol, and 0.1 M Tris-HCl, pH 8.5, X-ray diffraction structure determination and analysis at 1.3 A resolution, modeling of 2 enzyme molecules, residues 1-359, complexed with 2 molecules of NADP+, 2 ions of Fe2+, 2 molecules of Tris, and 895 water molecules

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6 - 10

purified recombinant His-tagged enzyme, 50% activityy remaining after 1 h and inactivation aafter 5 h at pH 6.0, 50% activity remaining after 3 h and 30% after 5 h at pH 7.5, inactivation after 1 h at pH 10.0

739886

6 - 7

most stable within this range

721395

additional information

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15 - 45

purified recombinant His-tagged enzyme, 50% activity remaining after 3 h and 30% after 5 h at 30-37°C, inactivation after 2 h at 45°C

60% of enzyme relative activity remain after a 2h incubation at 37°C

60 min, inactivation

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-20°C, Mn2+-DTT-HEPES buffer, stable for 3 days, 82% loss of activity after 3 weeks

389488

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inactivated by oxidation during aerobic metabolism

389488

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5 entries

by centrifugation, nickel affinity column and gel filtration

recombinant His-tagged enzyme 7.1-7.3fold from Escherichia coli strain JM109 by nickel affinity chromatography and gel filtration

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

recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration

recombinant protein

recombinant soluble enzyme from Escherichia coli strain BL21(DE3)

670216

recombinant tagged selenomethionine-labeled enzyme from Escherichia coli by nickel affinity, ion exchange, and gel filtration chromatography and ultrafiltration

Thermotoga maritima

Q9X022

657320

soluble recombinant His-tagged enzyme 6.6-6.7fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration to homogeneity

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all genes of the dha regulon, including the gene for 1,3-propanediol oxidoreductase of Klebsiella pneumoniae mobilized by the plasmid RP4:mini Mu and transferred to Escherichia coli

389493

expression in Clostridium acetobutylicum mutant strain DG1 from plasmid pSPD5, subcloning in Escherichia coli

667270

expression in Escherichia coli

gene dhaT, cloning from genomic DNA, DNA sequence determination, inducible high-level expression of the mostly soluble enzyme in Escherichia coli strain BL21(DE3)

670216

gene dhaT, DNA and amino acid sequence determination and analysis, sequence comparisons

A3RL85

688171

gene dhaT, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain JM109

gene dhaT, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in YqhD-deficient Escherichia coli strain BL21(DE3)

gene dhaT, expression of the N-terminally His-tagged in Escherichia coli strain BL21(DE3)

gene dhaT, high level co-expression of the 1,3-PD oxidoreductase with glycerol dehydratase, encoded by gene dhaB, and glycerol dehydratase reactivating factor, encoded by gene gdrAB, in Escherichia coli strain BL21 (DE3) using two incompatible plasmids, fed-batch fermentation of recombinant bacteria. The NADPH-linked alcohol dehydrogenase, which is encoded by yqhD, a gene from Escherichia coli, can non-specifically catalyze 3-hydroxypropionaldehyde converting to 1,3-propanediol with sufficient activity, overview

675901

gene dhaT, part of the dha regulon, genetic structure, overview. Coexpressions of the PDOR and GDHt from gene dhaB in Klebsiella pneumoniae result in an increase of molar yield from 50.6-64.0% of 1,3-propanediol

gene dhaT, part of the dha regulon, genetic structure, recombinant expression in Escherichia coli

gene dhaT, part of the dha regulon, recombinant expression in Escherichia coli

6 entries

gene dhaT, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3)

740927

gene TM0920, expression of soluble N-terminally MGSDKIHHHHHH-tagged selenomethionine-labeled enzyme in Escherichia coli methionine auxotrophic strain DL41

Thermotoga maritima

Q9X022

657320

genes for the production of propane-1,3-diol in Clostridium butyricum, dhaB1 and dhaB2, which encode the vitamin B12-independent glycerol dehydratase DhaB1 and its activating factor, DhaB2, respectively, tandemly arrayed with the Escherichia coli yqhD gene, which encodes the 1,3-propanediol oxidoreductase isoenzyme YqhD. Heterologous expression of the propane-1,3-diol operon under the control of the temperature-sensitive lambda phage PLPR promoter regulated by the cIts857 repressor, from plasmid pDY220, in Escherichia coli K-12 ER2925

Escherichia coli

695764

overexpression in Escherichia coli

389494

overexpression of gene dhaT encoding PDOR (from vector pETPkan-dhaT) and transformed into Klebsiella pneumoniae

695716

plasmid including Escherichia coli yqhD introduced into Klebsiella pneumoniae AK mutant strain defective in in 1,3-PD oxidoreductase activity (DhaT)

Escherichia coli

695844

plasmid pUC18K-dhaT transformed into Klebsiella pneumoniae KG1

699149

plasmids pUC18 K-dhaT and pUC18 K-gdh, carrying the genes dhaT encoding PDOR and gdh encoding glycerol dehydrogenase, respectively, transformed into a propane-1,3-diol native producer Klebsiella pneumoniae KG1

696785

recombinant plasmids containing dhaT expressed in Klebsiella pneumoniae Cu, and the mutant strains AK and AR

695842

the dhaD gene encoding glycerol dehydrogenase (GDH) and dhaT gene encoding 1,3-propanediol oxidoreductase (PDOR) inserted into pTD plasmid and overexpressed in Klebsiella pneumoniae ACCC 10082

697666

the dhaT gene coding for 1,3-PD dehydrogenase inserted into vector pET-YSBLIC and transformed into Escherichia coli BL21(DE3)

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nore

D41A

mutation for relaxation of the coenzyme specificity, weakens the repulsion between Asp41 and the phosphate group esterified to the 2-hydroxyl group of the ribose at the adenine end of NADPH

D41G

mutation for relaxation of the coenzyme specificity, weakens the repulsion between Asp41 and the phosphate group esterified to the 2-hydroxyl group of the ribose at the adenine end of NADPH