Information on EC 1.1.2.8 - alcohol dehydrogenase (cytochrome c)

-

Substrates: -
Products: -

?

1,2-cyclopentandiol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

1,3-butanediol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

1-butanol + phenazine methosulfate

butanal + reduced phenazine methosulfate

Substrates: -
Products: -

?

1-cyclohexylethanol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

1-hexanol + phenazine methosulfate

hexanal + reduced phenazine methosulfate

Substrates: -
Products: -

?

1-octanol + phenazine methosulfate

octanal + reduced phenazine methosulfate

Substrates: -
Products: -

?

2,3-butanediol + phenazine methosulfate

? + reduced phenazine methosulfate

2,4-pentanediol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

2-butanol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

2-butanol + phenazine methosulfate

butan-2-one + reduced phenazine methosulfate

Substrates: -
Products: -

?

2-hexanol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

2-phenylethanol + phenazine methosulfate

phenylacetaldehyde + reduced phenazine methosulfate

acetaldehyde + phenazine methosulfate

? + reduced phenazine methosulfate

butanol + 2 cytochrome c

butyraldehyde + 2 reduced cytochrome c

butyraldehyde + phenazine methosulfate

? + reduced phenazine methosulfate

Substrates: -
Products: -

?

cinnamyl alcohol + phenazine methosulfate

cinnamyl aldehyde + reduced phenazine methosulfate

Substrates: -
Products: -

?

cis-1,2-cyclohexanediol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

cyclohexanol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

D-arabinose + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

D-arabitol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

D-galactose + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

D-gluconate + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

D-lyxose + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

D-mannitol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

D-ribose + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

D-sorbitol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

D-xylose + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

diethylstilbestrol + pyrroloquinoline quinone

diethylstilbestrol 4-semiquinone + pyrroloquinoline quinol

ethanol + 2 cytochrome c

acetaldehyde + 2 reduced cytochrome c

ethanol + 2 cytochrome c

ethanal + 2 reduced cytochrome c

ethanol + 2 ferricytochrome c

ethanal + 2 ferrocytochrome c

ethanol + N,N,N',N'-tetramethyl-p-phenylenediamine

ethanal + ?

-

Substrates: i.e. Wurster's Blue
Products: -

?

ethanol + oxidized 2,6-dichlorophenolindophenol

ethanal + reduced 2,6-dichlorophenolindophenol

ethanol + phenazine methosulfate

acetaldehyde + reduced phenazine methosulfate

763436, 763417

Substrates: -
Products: -

?

ethanolamine + phenazine methosulfate

? + reduced phenazine methosulfate

Substrates: -
Products: -

?

farnesol + phenazine methosulfate

farnesal + reduced phenazine methosulfate

Substrates: -
Products: -

?

glyceraldehyde + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

glycerol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

hexanal + phenazine methosulfate

? + reduced phenazine methosulfate

Substrates: -
Products: -

?

isopropanol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

L-arabinose + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

L-ribose + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

meso-erythritol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

methanol + 2 cytochrome c

formaldehyde + 2 reduced cytochrome c

methanol + phenazine methosulfate

formaldehyde + reduced phenazine methosulfate

Substrates: -
Products: -

?

octanal + phenazine methosulfate

? + reduced phenazine methosulfate

Substrates: -
Products: -

?

propanol + 2 cytochrome c

propanal + 2 reduced cytochrome c

ribitol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

threitol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

trans-1,2-cyclohexanediol + phenazine methosulfate

? + reduced phenazine methosulfate

-

Substrates: -
Products: -

?

additional information

?

-

5 entries

butanol + 2 cytochrome c

butyraldehyde + 2 reduced cytochrome c

diethylstilbestrol + pyrroloquinoline quinone

diethylstilbestrol 4-semiquinone + pyrroloquinoline quinol

ethanol + 2 cytochrome c

acetaldehyde + 2 reduced cytochrome c

ethanol + 2 ferricytochrome c

ethanal + 2 ferrocytochrome c

methanol + 2 cytochrome c

formaldehyde + 2 reduced cytochrome c

additional information

?

-

cytochrome c

pyrroloquinoline quinone

13 entries

additional information

-

Ca2+

Iron

-

one [2Fe-2S] cluster per enzyme

La3+

-

required, active site-bound, 1.3 mol of La3+ per mol of ExaF protomer, indicating a 1:1 ratio of L3+ to protomer of enzyme

Mg2+

-

required

Sr2+

-

incubation of apo-enzyme with Sr2+ and pyrroloquinoline quinone leads to the formation of an active Sr2+-form. The Sr2+ and the Ca2+-forms of the enzyme differ in their absorption spectra. The Sr2+-form is inactivated by trans-l,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid twice as fast as the Ca2+-form.

701600

additional information

-

the enzyme from strain AM1 utilizes La3+ rather than Ca2+ as a cofactor

ethylamine

-

activating at low concentrations, inhibitory at high concentrations

pentylamine

-

the alcohol oxidation activity of PpADH is strongly enhanced by pentylamine and the affinity for substrates is also improved by pentylamine as an activator, inhibitory at high concentrations

ammonium

-

Butylamine

-

Dimethylamine

-

dipropylamine

-

ethanol

-

does not affect the adhS gene expression but induces PQQ-ADH activity

ethylamine

-

inhibitory at high concentrations

heptylamine

-

Hexylamine

-

Isobutylamine

-

slight activation

methylamine

octylamine

-

pentylamine

-

the alcohol oxidation activity of PpADH is strongly enhanced by pentylamine and the affinity for substrates is also improved by pentylamine as an activator, inhibitory at high concentrations

Propylamine

-

triethylamine

-

trimethylamine

-

tripropylamine

-

additional information

-

the enzyme requires ammonia or primary amines as activators in in vitro assays with artificial electron acceptors, the enzyme is activated by various primary amines, dimethylamine, and trimethylamine. tert-Butylamine is a poor activator. The hydrophobic interaction contributes to the binding of amine activators to the binding site in the enzyme

-

0.115

2-phenylethanol

pH 8.0, 30°C

0.899

acetaldehyde

pH 8.0, 30°C

0.061 - 0.21

ethanol

5 entries

0.35 - 0.43

Propanol

24.8 - 56.6

ethanol

32.6 - 73.1

Propanol

0.051

2-phenylethanol

pH 8.0, 30°C

0.007

acetaldehyde

pH 8.0, 30°C

0.081 - 395

ethanol

5 entries

73 - 209

Propanol

112.5

ethylamine

-

pH 9.0, temperature not specified in the publication

37.6

pentylamine

-

pH 9.0, temperature not specified in the publication

0.0031

-

substrate: threitol, pH 5.0, 25°C

0.0052

-

substrate: D-arabinose, pH 5.0, 25°C

0.01

-

substrate: trans-1,2-cyclohexanediol, pH 5.0, 25°C

0.018

-

substrate: glycerol, pH 5.0, 25°C

0.022

-

substrate: 1-cyclohexylethanol, pH 5.0, 25°C

0.027

-

substrate: D-mannitol, pH 5.0, 25°C

0.028

-

substrate: ribitol, pH 5.0, 25°C

0.03

-

substrate: 2-hexanol, pH 5.0, 25°C

0.035

-

substrate: isopropanol, pH 5.0, 25°C

0.055

-

substrate: D-sorbitol, pH 5.0, 25°C

0.065

-

substrate: 1,3-butanediol, pH 5.0, 25°C

0.08

-

substrate: 1,2-butanediol, pH 5.0, 25°C

0.084

-

substrate: cyclohexanol, pH 5.0, 25°C

0.085

-

substrate: 2,4-pentanediol, pH 5.0, 25°C

0.09

-

substrate: D-ribose, pH 5.0, 25°C

0.1

-

substrate: D-lyxose, pH 5.0, 25°C

0.12

0.13

0.14

0.17

-

substrate: D-arabitol, pH 5.0, 25°C

0.19

-

substrate: meso-erythritol, pH 5.0, 25°C

0.2

-

substrate: L-ribose, pH 5.0, 25°C

0.35

-

substrate: 1,2-cyclopentandiol, pH 5.0, 25°C

0.4

-

substrate: 2,3-butanediol, pH 5.0, 25°C

0.55

-

substrate: cis-1,2-cyclohexanediol, pH 5.0, 25°C

40

-

pH 9.0, 25°C

6

C7G3B7, D2SZY4, C9K501, C7G3B8, D2SZY5, C9W9E3, C9K502, C9K504

assay at

7.5

-

assay at

9

25

30

35

C7G3B7, D2SZY4, C9K501, C7G3B8, D2SZY5, C9W9E3, C9K502, C9K504

-

Ectopseudomonas mendocina

6.77

-

sequence calculation

6.96

Stutzerimonas stutzeri

-

sequence calculation

Acetobacter pasteurianus IFO3191

10 entries

Acetobacter pasteurianus MSU10

Acetobacter pasteurianus JST-S

KY643658: subunits I (adhA), KY584296: subunit II (adhB)

763081

KY643658; KY584296

GenBank

Azospirillum brasilense Sp7

Azospirillum brasilense

-

763204

-

-

763204

-

Ectopseudomonas mendocina

-

Ectopseudomonas mendocina YMP

-

-

-

-

Gluconacetobacter diazotrophicus PAL5

-

-

-

-

-

-

-

Methylorubrum extorquens ATCC 14718 / DSM 1338 / JCM 2805 / NCIMB 9133 / AM1

-

-

Pseudomonas putida KT 2240

-

-

-

-

Stutzerimonas stutzeri

-

-

Stutzerimonas stutzeri ATCC 14405

-

-

additional information

Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

evolution

8 entries

malfunction

metabolism

physiological function

13 entries

additional information

Methylacidiphilum fumariolicum SolV

Pseudogluconobacter saccharoketogenes

Pseudomonas putida KT2440

115000

-

inactive enzyme form, native PAGE

120000

-

inactive enzyme form, gel filtration

126400

-

recombinant His6-tagged enzyme, gel filtration

136000

-

gel filtration

330000

-

active enzyme form, gel filtration

345000

-

active enzyme form, native PAGE

45000

60000

72000

9000

?

dimer

1 * 60000 + 1 * 9000, SDS-PAGE

heterodimer

heterotetramer

homodimer

tetramer

-

2 * 60000, alpha-subunit, + 2 * 9000, beta-subunit, SDS-PAGE

additional information

-

enzyme interacts with a soluble cytochrome cEDH, the oxidized form being an excellent acceptor for the semiquinone form of EDH. This cytochrome is quite different from the cytochrome c551 operating in nitrate respiration

additional information

sequence contains a signal peptide of 34 residues

389964

alignment with the amino acid sequence of the large subunit of the quinoprotein methanol dehydrogenase from Methylobacterium extorquens. The amino acid residues involved in the binding of pyrroloquinoline quinone and Ca2+ at the active site are conserved

389964

diffraction to beyond 2.5 A, space group R3

-

705106

to 2.6 A resolution, by molecular replacement. Eight W-shaped beta-sheet motifs are arranged circularly in a propeller-like fashion forming a disk-shaped superbarrel. The prosthetic group is located in the centre of the superbarrel and is coordinated to a calcium ion. Most amino acid residues found in close contact with the prosthetic group pyrroloquinoline quinone and the Ca2+ are conserved between the quinoprotein ethanol dehydrogenase structure and that of the methanol dehydrogenases from Methylobacterium extorquens or Methylophilus W3A1. The main differences in the active-site region are a bulky tryptophan residue in the active-site cavity of methanol dehydrogenase, which is replaced by a phenylalanine and a leucine side-chain in the ethanol dehydrogenase structure and a leucine residue right above the pyrrolquinoline quinone group in methanol dehydrogenase which is replaced by a tryptophan side-chain. Both amino acid exchanges contribute to different substrate specificities of these otherwise very similar enzymes. In addition to the Ca2+ in the active-site cavity, ethanol dehydrogenase contains a second Ca2+-binding site at the N-terminus, which contributes to the stability of the native enzyme

-

389959

A26V

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site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

G55D

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

L18Q

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

T104K

-

site-directed mutagenesis, the mutation leads to a complete loss of ethanol oxidizing ability

V107A

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

V36I

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

V54I

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

V70A

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

A26V

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

-

G55D

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

-

L18Q

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

-

V36I

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

-

V54I

-

site-directed mutagenesis, the mutation does not affect the PQQ-ADH activity and ethanol oxidizing ability

-

C105A/C106A

-

mutation of residues forming a characteristic disulfide ring in the binding pocket of pyrroloquinoline quinone. Analysis by EPR spectroscopy shows that the disulfide ring is no prerequisite for the formation of the functionally important semiquinone form of pyrroloquinoline quinone

674560

E408P

mutant shows a 2.3fold increased stability upon incubation at 45°C for 1 h compared with the wild-type allele

N410K

44% relative residual activities compared with 23% for the wild-type allele upon 1 h incubation at 45°C

N410S

about 30% relative residual activities compared with 23% for the wild-type allele upon 1 h incubation at 45°C

N410T

about 35% relative residual activities compared with 23% for the wild-type allele upon 1 h incubation at 45°C

R91D

41% relative residual activities compared with 23% for the wild-type allele upon 1 h incubation at 45°C

R91D/E408P

mutant shows a 3.2fold stability upon incubation at 45°C for 1 h compared with the wild-type allele

R91D/E408P/N410K

R91E

about 35% relative residual activities compared with 23% for the wild-type allele upon 1 h incubation at 45°C

R91Q

about 30% relative residual activities compared with 23% for the wild-type allele upon 1 h incubation at 45°C

additional information

10 entries

45

relative residual activity upon 1 h incubation: about 20% (wild-type enzyme), 41% (mutant enzyme R19D), about 35% (mutant enzyme R191E), about 30% (mutant enzyme R191Q), 44% (mutant enzyme N410K), about 40% (mutant enzyme N410T), about 30% (mutant enzyme N410S), 69% (mutant enzyme E408P)

acetic acid

-

-

C-terminally His-tagged PQQ-ADHs are expressed in Escherichia coli BL21(DE3)

native enzyme from membranes by two steps of anion exchange chormatography, hydroxyapatite chromatography, and gel filtration

-

native enzyme from strain IFO3191 by ultracentrifugation and anion exchange chromatography

C7G3B7, D2SZY4, C9K501, C7G3B8, D2SZY5, C9W9E3, C9K502, C9K504

native enzyme from strain MSU10 by ultracentrifugation and anion exchange chromatography

C7G3B7, D2SZY4, C9K501, C7G3B8, D2SZY5, C9W9E3, C9K502, C9K504

native enzyme from strain SKU1108 by ultracentrifugation and anion exchange chromatography

C7G3B7, D2SZY4, C9K501, C7G3B8, D2SZY5, C9W9E3, C9K502, C9K504

recombinant His6-tagged enzyme from Escherichia coli strain TOP10 by ultracentrifugation, nickel affinity chromatography, ultrafiltration, and desalting gel filtration

-

-

389964

a plasmid construct is used to express PQQ-DH9. DH9. The expression host is a derivative Gluconobacter sp. strain of CHM43, which lacks the genes for PQQ-dependent glycerol dehydrogenase and the membrane-bound alcohol dehydrogenase and consequently has minimal ability to oxidize primary and secondary alcohols. The membranes of the transformant exhibited considerable d-arabitol dehydrogenase activity, whereas the reference strain did not, even if it had PQQ-DH9-encoding genes in the chromosome and harbored the empty vector. This suggests that PQQ-DH9 is not expressed in the genome

-

Acetobacter pasteurianus JSTS subunits I (adhA) and II (adhB) of the enzyme (PQQ-ADH) are amplified by PCR, subcloned into a vector, and transformed back into Acetobacter pasteurianus JST-S. The acetic acid production is improved by the engineered strain (61.42 g/l) while the residual ethanol content (4.18 g/l) is decreased

KY643658; KY584296

763081

C-terminally His-tagged wild-type and mutant enzyme forms are expressed in Escherichia coli BL21(DE3)

gene adhA, DNA and amino acid sequence determination and analysis, sequence comparison of the genes encoding subunit I, subunit II, and subunit III of ADHs

C7G3B7, D2SZY4, C9K501, C7G3B8, D2SZY5, C9W9E3, C9K502, C9K504

gene adhB, DNA and amino acid sequence determination and analysis, sequence comparison of the genes encoding subunit I, subunit II, and subunit III of ADHs

C7G3B7, D2SZY4, C9K501, C7G3B8, D2SZY5, C9W9E3, C9K502, C9K504

gene adhS, DNA and amino acid sequence determination and analysis, sequence comparison of the genes encoding subunit I, subunit II, and subunit III of ADHs

C7G3B7, D2SZY4, C9K501, C7G3B8, D2SZY5, C9W9E3, C9K502, C9K504

gene exaF, or exaA, DNA and amino acid sequence determination and analysis, functional recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain TOP10

-

sequence comparisons and phylogenetic tree, real-time reverse transcriptase PCR expression analysis

ethanol does not affect the adhS gene expression but induces PQQ-ADH activity

expression is upregulated when Azospirillum brasilense Sp7 is grown in medium containing glycerol as a source of carbon

the enzyme is induced by diethylstilbestrol, presence of diethylstilbestrol causes a 5.2fold increase in the mRNA level of the gene encoding quinoprotein alcohol dehydrogenase

treatment with trans-l,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid at 30°C leads to an catalytically inactive apo-form. Upon incubation of the apo-form with Ca2+ and pyrroloquinoline quinone a fully active holo-enzyme is reconstituted. Incubation of apo-enzyme with Sr2+ and pyrroloquinoline quinone leads to the formation of an active Sr2+-form. The Sr2+ and the Ca2+-forms of the enzyme differ in their absorption spectra.

-

701600

environmental protection