Information on EC 1.2.7.4 - anaerobic carbon-monoxide dehydrogenase

-

690057

CODHII

coos

11 entries

Dehydrogenase, carbon monoxide

-

Ni,Fe-CODH

-

712750

Ni,Fe-dependent carbon monoxide dehydrogenase

Desulfotomaculum nigrificans

-

712750

Ni-CODH

-

740945

Ni-containing carbon monoxide dehydrogenase

Desulfotomaculum nigrificans

-

740945

Ni-Fe carbon monoxide dehydrogenase II

CO + H2O + 2 oxidized ferredoxin = CO2 + 2 reduced ferredoxin + 2 H+

reductive acetyl coenzyme A pathway

oxidation

-

redox reaction

-

reduction

-

BRENDA

KEGG

Carbon fixation pathways in prokaryotes, Methane metabolism, Microbial metabolism in diverse environments, Nitrotoluene degradation

MetaCyc

carbon tetrachloride degradation II, hydrogen production VI, methanogenesis from acetate, reductive acetyl coenzyme A pathway I (homoacetogenic bacteria), reductive acetyl coenzyme A pathway II (autotrophic methanogens)

carbon-monoxide,water:ferredoxin oxidoreductase

This prokaryotic enzyme catalyses the reversible reduction of CO2 to CO. The electrons are transferred to redox proteins such as ferredoxin. In purple sulfur bacteria and methanogenic archaea it catalyses the oxidation of CO to CO2, which is incorporated by the Calvin-Benson-Basham cycle or released, respectively. In acetogenic and sulfate-reducing microbes it catalyses the reduction of CO2 to CO, which is incorporated into acetyl CoA by EC 2.3.1.169, CO-methylating acetyl CoA synthase, with which the enzyme forms a tight complex in those organisms. The enzyme contains five metal clusters per homodimeric enzyme: two nickel-iron-sulfur clusters called the C-Clusters, one [4Fe-4S] D-cluster; and two [4Fe-4S] B-clusters. In methanogenic archaea additional [4Fe-4S] clusters exist, presumably as part of the electron transfer chain. In purple sulfur bacteria the enzyme forms complexes with the Ni-Fe-S protein EC 1.12.7.2, ferredoxin hydrogenase, which catalyse the overall reaction: CO + H2O = CO2 + H2. cf. EC 1.2.5.3, aerobic carbon monoxide dehydrogenase.

64972-88-9

-

acetyl-SCoA + CO

-

acetyl-CoA/CO exchange reaction, 14C experiments prove that the enzyme can cleave both the carbon-carbon and carbon-sulfur bonds of acetyl-CoA as well as to store methyl, CO, and CoA fragments at the active site

-

r

acetyl-SCoA + CoASH

-

acetyl-CoA/CoA exchange reaction

-

r

CO + ferredoxin

?

-

ferredoxin is the direct electron acceptor, the carbon-monoxide dehydrogenase complex reduces a ferredoxin which, together with membranes and associated hydrogenase, reconstitutes a CO-oxidizing:H2-evolving system

390472

-

?

CO + H2O

CO2 + H2

-

-

?

CO + H2O + 1,1'-trimethylene-4,4'-dimethyl-2,2'dipyridylium dibromide

CO2 + reduced 1,1'-trimethylene-4,4'-dimethyl-2,2'dipyridylium dibromide

-

-

?

CO + H2O + 2,3,5-triphenyltetrazolium chloride

CO2 + reduced 2,3,5-triphenyltetrazolium chloride

CO + H2O + 2-(4-iodophenyl)-3-(4-nitrophenyl)-2H-tetrazolium chloride

CO2 + reduced 2-(4-iodophenyl)-3-(4-nitrophenyl)-2H-tetrazolium chloride

CO + H2O + acceptor

CO2 + reduced acceptor

27 entries

CO + H2O + acceptor

CO2 + reduced acceptor + H+

-

-

?

CO + H2O + benzyl viologen

CO2 + reduced benzyl viologen

-

-

?

CO + H2O + coenzyme F420

CO2 + reduced coenzyme F420

CO + H2O + cytochrome b

CO2 + reduced cytochrome b

-

-

?

CO + H2O + cytochrome c3

CO2 + reduced cytochrome c3

-

cytochrome 3 from Desulfovibrio vulgaris

-

?

CO + H2O + electron acceptor

?

15 entries

CO + H2O + electron acceptor

CO2 + reduced electron acceptor

-

a proton gradient across the cytoplasmic membrane is generated by channeling the electrons formed via cytochrome b561 into a CO-insensitive respiratory chain

-

?

CO + H2O + FAD

CO2 + FADH2

CO + H2O + ferredoxin

CO2 + reduced ferredoxin

CO + H2O + flavodoxin

CO2 + reduced flavodoxin

-

-

?

CO + H2O + FMN

CO2 + FAD

CO + H2O + FMN

CO2 + FMNH2

7 entries

CO + H2O + methyl viologen

CO2 + reduced methyl viologen

33 entries

CO + H2O + methyl viologen

CO2 + reduced methyl viologen + H+

CO + H2O + methylene blue

CO2 + reduced methylene blue

CO + H2O + methylene blue

CO2 + reduced methylene blue + H2

-

-

?

CO + H2O + NAD+

CO2 + NADH + H+

CO + H2O + NADP+

CO2 + NADPH + H+

CO + H2O + oxidized 2,3,5-triphenyltetrazolium chloride

CO2 + reduced 2,3,5-triphenyltetrazolium chloride

-

-

?

CO + H2O + oxidized 2-(4-iodophenyl)-3-(4-nitrophenyl)-2H-tetrazolium chloride

CO2 + reduced 2-(4-iodophenyl)-3-(4-nitrophenyl)-2H-tetrazolium chloride

CO + H2O + oxidized cytochrome b

CO2 + reduced cytochrome b

-

membrane-bound b-type, native electron carrier

-

?

CO + H2O + oxidized cytochrome c3

CO2 + reduced cytochrome c3

-

Desulfovibrio vulgaris cytochrome c3

-

?

CO + H2O + oxidized ferredoxin

CO2 + reduced ferredoxin

7 entries

CO + H2O + oxidized ferredoxin

CO2 + reduced ferredoxin + H+

CO + H2O + oxidized flavodoxin

CO2 + reduced flavodoxin

CO + H2O + oxidized methyl viologen

CO2 + reduced methyl viologen

CO + H2O + oxidized methyl viologen

CO2 + reduced methyl viologen + H+

CO + H2O + oxidized methylene blue

CO2 + reduced methylene blue

-

-

?

CO + H2O + oxidized phenazine methosulfate

CO2 + reduced phenazine methosulfate

-

-

?

CO + H2O + oxidized rubredoxin

CO2 + reduced rubredoxin

-

most efficient electron acceptor

-

?

CO + H2O + oxidized viologen

CO2 + reduced viologen

-

-

?

CO + H2O + phenazine methosulfate

CO2 + reduced phenazine methosulfate

-

-

?

CO + H2O + rubredoxin

CO2 + reduced rubredoxin

H2 + reduced methyl viologen

methyl viologen

-

-

r

propionyl-SCoA + CoASH

-

acetyl-CoA/CoA exchange reaction

-

r

tetrahydrosarcinapterin + acetyl-SCoA + H2O

methyltetrahydrosarcinapterin + CO2 + CoASH

trichloroethylene + ?

cis-dichloroethylene + trans-dichlororethylene + 1,1-dichloroethylene + vinyl chloride + ethylene

-

-

?

additional information

?

-

CO + ferredoxin

?

-

ferredoxin is the direct electron acceptor, the carbon-monoxide dehydrogenase complex reduces a ferredoxin which, together with membranes and associated hydrogenase, reconstitutes a CO-oxidizing:H2-evolving system

390472

-

?

CO + H2O

CO2 + H2

-

-

?

CO + H2O + acceptor

CO2 + reduced acceptor

11 entries

CO + H2O + acceptor

CO2 + reduced acceptor + H+

-

-

?

CO + H2O + electron acceptor

?

15 entries

CO + H2O + electron acceptor

CO2 + reduced electron acceptor

-

a proton gradient across the cytoplasmic membrane is generated by channeling the electrons formed via cytochrome b561 into a CO-insensitive respiratory chain

-

?

CO + H2O + oxidized cytochrome b

CO2 + reduced cytochrome b

-

membrane-bound b-type, native electron carrier

-

?

CO + H2O + oxidized ferredoxin

CO2 + reduced ferredoxin

CO + H2O + oxidized ferredoxin

CO2 + reduced ferredoxin + H+

additional information

?

-

-

Thermacetogenium phaeum operates the CO dehydrogenase/acetyl-CoA pathway reversibly both in acetate oxidation and in reductive acetogenesis by using the same biochemical apparatus

-

FAD

Fe-S center

-

flavin adenine dinucleotide

iron-sulfur centre

-

712300

molybdopterin

-

657190

molybdopterin cytosine dinucleotide

Ni-4Fe-5S center

-

active site cluster

Ni-Fe-4S center

-

active site called the C-cluster

687824

Ni-Fe-S center

-

uses a Ni-Fe-S center called the C-cluster to reduce carbon dioxide to carbon monoxide and uses a second Ni-Fe-S center, called the A-cluster, to assemble acetyl-CoA from a methyl group, coenzyme A, and C-cluster-generated CO

-

4Fe-4S cluster

-

core extrusion experiments indicate 6 [4Fe-4S] clusters per tetramer, and electron paramagnetic resonance spectroscopy detects at least one of these clusters, in the reduced form

727806

Co2+

-

the multifunctional enzyme complex contains Co2+

copper

Cu2+

Fe-S center

-

the multifunctional enzyme complex contains iron-sulfur centers

Fe2+

Iron

Molybdenum

Ni-Fe-S center

-

Ni2+

16 entries

Nickel

21 entries

S2-

-

contains a [NiFe4S5] center called cluster C

690057

Zinc

Zn2+

-

the multifunctional enzyme complex contains Zn2+

[2Fe-2S]-center

-

the small subunit contains two [2Fe-2S] centers

725197

additional information

1,10-phenanthroline

-

1 mM, inactivates CO/acetyl-CoA exchange activity completely but has no effect on CO oxidation

390462

2,3-Butanedione

-

5,5'-dithiobis (2-nitrobenzoate)

-

390459

butyryl-CoA

-

CN-

10 entries

CO

CoASH

-

390468, 390469

COS

-

rapid-equilibrium inhibitor largely competitive versus CO, uncompetitive versus methyl viologen

390466

crotonyl-CoA

-

cyanide

D-glucose

Labrenzia aggregata

Q7X0G3

partially inhibited by incubation with 0.5 mM exogenous D-glucose

dephospho-CoASH

-

desulfo-CoA

-

Glyoxaldehyde

-

inactivation requires enzymatic turnover; only with CO and methyl viologen as substrates

Hg+

-

390456

KCN

Mersalyl acid

-

390459

methyl iodide

N2O

-

inhibition of the exchange reaction between acetyl-CoA and CoA

390475

O2

PCMB

-

390456

pivaloylpantetheine-SH

-

potassium cyanide

-

competitive inhibitor of reduced CODHII with respect to the substrate CO, inhibition of dithionite- or Ti(III) citrate-reduced CODHII by potassium cyanide is fully reversible since the enzyme can be completely reactivated, sodium sulfide has no effect on the reactivation of cyanide-inhibited CODHII in the presence of dithionite

propionyl-CoA

-

S2-

SCN-

-

binds to C-center, mixed partial inhibition

390461

Sulfide

thionin

-

654737

additional information

-

corrinoid cofactor

-

the multifunctional enzyme complex contains a corrinoid cofactor

-

Nickel

-

Ni-activated alpha subunit rapidly and reversibly accepts a methyl group

sodium nitroprusside

Ti(III)citrate

-

1mM for 1 min at 65°C, 45fold increase in initial activity

additional information

-

CooC proteins are ATPases involved in the incorporation of nickel into the complex active site ([Ni-4Fe-4S]) cluster of Ni,Fe-dependent carbon monoxide dehydrogenases

712750

-

0.5

2,3,5-triphenyltetrazolium chloride

-

below 0.5 mM; pH 7.0, 25°C

0.6 - 1.5

acetyl-CoA

0.01 - 5

CO

0.05

CoA

-

acetyl-CoA/CoA exchange reaction

0.008

Ferredoxin

-

390481

-

0.065 - 7.1

methyl viologen

0.00295 - 0.0107

methylene blue

5

propionyl-CoA

-

1.7

Trichloroethylene

-

1.17 - 15900

CO

8.2 - 93.3

methylene blue

additional information

-

2780 - 8700

methylene blue

1.45

butyryl-CoA

-

inhbition of CO/acetyl-CoA exchange reaction

0.4 - 1.5

CO

0.007

CoASH

0.0023 - 0.0158

COS

1.4

crotonyl-CoA

-

inhbition of CO/acetyl-CoA exchange reaction

0.0217

cyanide

-

at 23°C, in 4 mM dithionite, and 4 mM dithiothreitol

0.035

dephospho-CoASH

-

inhbition of CO/acetyl-CoA exchange reaction

6

desulfo-CoA

-

inhbition of CO/acetyl-CoA exchange reaction

1.2

pivaloylpantetheine-SH

-

inhbition of CO/acetyl-CoA exchange reaction

1.3

propionyl-CoA

-

inhbition of CO/acetyl-CoA exchange reaction

16 - 830

SCN

0.00004 - 0.00006

Sulfide

0.001 - 0.002

-

CO2/acetyl-CoA exchange in cell extracts at 85°C

390471

0.006

-

at 37°C

Mycobacterium tuberculosis

0.0099

-

at 37°C

0.0175

Mycolicibacterium vaccae

-

at 37°C

Archaeoglobus lithotrophicus

0.04

-

pH 7.5, 65°C, cell extract

0.0577

-

0.1

Ferroglobus placidus

-

pH 7.5, 65°C

721443

0.32

-

CO/acetyl-CoA exchange reaction

390462

0.4

0.9

-

pH 8.1, 80°C

723991

0.96

-

after 12.92fold purification, at 95°C, pH 8.0, using methylene blue as electron acceptor

2.08

-

after 12.92fold purification, at 95°C, pH 8.0, using methyl viologen as electron acceptor

2.45

-

after 12.92fold purification, at 95°C, pH 8.0, using NADP+ as electron acceptor

2.47

-

after 12.92fold purification, at 95°C, pH 8.0, using NAD+ as electron acceptor

5.2

-

pH 7.5, 65°C, cell extract

5.7

-

CO oxidation in cell-extracts at 85°C, electron acceptor methylviologen

390471

6.9

-

mutant C531A, pH 7.5

7

-

acetyl-CoA/CoA exchange reaction at pH 6.0 and 45°C, 100% CO in the gas phase

390475

28

-

acetyl-CoA/CoA exchange reaction at pH 6.0 and 45°C, CO in the gas phase replaced by N2

390475

33

-

40

-

390465

76.8

-

cells grown in pure culture on pyruvate, pH 7.5, 55°C

86.3

-

mutant C451S, pH 7.5

100

-

25°C, mutant A265M

117

-

120

-

electron acceptor ferredoxin

390451

133.6

-

163

-

cells grown syntrophically on acetate, pH 7.5, 55°C

190

-

200

-

25°C, wild type

216

-

pH 7.0, 37°C

220

-

250

-

CO oxidation

390462

280

-

25°C, mutant A110C

300

-

25°C, mutant A222L

320

-

435

-

390453

675

-

electron acceptor methyl viologen

694

-

5924

-

pH 7.0, 70°C

6230

-

wild-type, pH 7.5

12000

-

390486

14000

-

carbon monoxide dehydrogenase II

additional information

5.3

-

390459

6.5

-

both for enzyme and complex

6.7

-

in cell-free extracts

390456

7 - 9

7.2

-

7.5

8 - 9

-

8.2 - 8.4

-

8.4

8.6

-

wild-type

9

10.5

-

10.6

-

; metyl viologen as substrate

5 - 11

-

very little activity outside this range

5 - 10

-

6.5 - 9

-

pH 6.5: about 25% of maximal activity, pH 8-9: maximal activity

8 - 11.5

-

pH 8: about 55% of maximal activity, pH 11.5: about 50% of maximal activity

8

-

about 60% of activity at pH 9.0

23

-

assay at

65

70

-

80

-

assay at

723991

90

-

95

-

75 - 100

-

about 65% activity at 75°C, about 75% activity at 80°C, about 80% activity at 85°C, about 90% activity at 90°C, 100% activity at 95°C, about 30% activity at 100°C, respectively

Acetobacterium woodii

-

-

-

-

-

-

Archaeoglobus fulgidus 7324

-

390471, 721441, 723991

-

-

723991

-

Archaeoglobus lithotrophicus

-

Archaeoglobus lithotrophicus TF-2

-

-

-

Autotrophic methanogenic bacterium

-

-

gram-negativ soil bacterium

-

Carboxydothermus hydrogenoformans DSM 6008

-

724590

Q9F8A8

Uniprot

Carboxydothermus hydrogenoformans Z-2901 / DSM 6008

strain Z-2901 (DSM 6008)

Citrobacter amalonaticus Y19

-

Citrobacter amalonaticus

-

740543

-

-

740543

-

Clostridium pasteurianum

-

390451

-

Desulfotomaculum nigrificans

-

740945

-

Ferroglobus placidus

chemolithoautotrophic

721443

-

Ferroglobus placidus DSM 10642

chemolithoautotrophic

721443

-

-

-

Labrenzia aggregata

large subunit, fragment; synonym Stappia aggregata

Methanococcus maripaludis

Q7X0G3

UniProt

more than threefold induction of enzyme activity and increase in enzyme expression in a mutant lacking the membrane-bound energy-conserving hydrogenase Ehb

674264

-

-

-

Methanosarcina barkeri DSM 800

-

390449, 390458, 390465, 390474, 727713, 727806

-

-

727806

-

strain Gö1

-

Methanosarcina thermophila TM-1

-

-

-

390456, 390458

-

-

-

Mycobacterium sp. DSM 3803 / JC1

-

Mycobacterium sp. JC1 DSM 3803

-

Mycobacterium tuberculosis

-

-

Mycobacterium tuberculosis H37Ra / ATCC 25177

-

-

Mycobacterium tuberculosis TMC 326 / H37Ra-INH-R / ATCC 35835

-

-

Mycolicibacterium vaccae

strain ATCC 15483

-

no activity in Methanobrevibacter ruminantium

-

-

no activity in Methanobrevibacter smithii

-

-

no activity in Methanococcus voltae

-

-

no activity in Methanospirillum hungatei

-

-

Stappia carboxidovorans

Stappia conradae

large subunit, fragment

A1E2F7

UniProt

Stappia kahanamokuae

large subunit, fragment

UniProt

-

-

large subunit, fragment

Q7X0G8

UniProt

Stappia sp. KB812

large subunit, fragment; strain KB812

Q7X0G9

UniProt

Stappia sp. KB902

large subunit, fragment

Q7X0G8

UniProt

Stappia sp. M8

large subunit, fragment; strain M8

Q7X0G5

UniProt

Stappia stellulata

large subunit, fragment

UniProt

grows on acetate if cocultured with hydrogen-consuming methanogenic partner Methanothermobacter thermautrophicus

Treponema azotonutricium ZAS-9

-

Treponema azotonutricium

-

726272

F5YDA7

UniProt

-

UniProt

-

UniProt

Treponema primitia ZAS-1

-

726272

F8TH35

UniProt

Treponema primitia ZAS-2

-

UniProt

-

F8TH37, F8TH38, F8TH39

UniProt

-

UniProt

-

UniProt

-

UniProt

culture condition:acetate-grown cell

culture condition:ammonium malate-grown cell

culture condition:formate-grown cell

-

culture condition:H2/sulfate-grown cell

culture condition:methanol-grown cell

-

5739

physiological function

Carboxydothermus hydrogenoformans (strain ATCC BAA-161 / DSM 6008 / Z-2901)

Desulfovibrio vulgaris (strain Hildenborough / ATCC 29579 / DSM 644 / NCIMB 8303)

3cf4, download, 3D-view

SCOPe (3cf4)

CATH (3cf4)

Q46G04

Methanosarcina barkeri (strain Fusaro / DSM 804)

1jqk, download, 3D-view

SCOPe (1jqk)

CATH (1jqk)

P31896

12600

-

1 * 86600 + 2 * 34500 + 1 * 12600, SDS-PAGE

17200

-

L2,M2,S2, 2 * 75000 + 2 * 28400 + 2 * 17200, SDS-PAGE

17390

D5G1Y2; D5G1Y0; D5G1Y1

small subunit, calculated from sequence of cDNA

712970

17752

-

L2,M2,S2, 2 * 87224 + 2 * 30694 + 2 * 17752, deduced from gene sequence, SDS-PAGE

17800

-

L2,M2,S2, 2 * 17800 + 2 * 30200 + 2 * 88700, dimer of heterotrimers, SDS-PAGE, crystal structure

18000

-

x * 92000 + x * 18000

19000

19400

-

2 * 19400 + 2 * 79400, SDS-PAGE

19700

21000

-

2 * 21000 + 2 * 89000, SDS-PAGE

28400

-

L2,M2,S2, 2 * 75000 + 2 * 28400 + 2 * 17200, SDS-PAGE

30200

-

L2,M2,S2, 2 * 17800 + 2 * 30200 + 2 * 88700, dimer of heterotrimers, SDS-PAGE, crystal structure

30620

D5G1Y2; D5G1Y0; D5G1Y1

medium subunit, calculated from sequence of cDNA

712970

30694

-

L2,M2,S2, 2 * 87224 + 2 * 30694 + 2 * 17752, deduced from gene sequence, SDS-PAGE

34500

-

1 * 86600 + 2 * 34500 + 1 * 12600, SDS-PAGE

50000

-

2 * 50000 + 2 * 71000 + 2 * 78000, alpha2beta2gamma2, SDS-PAGE

51400

-

6 * 19700, alpha, + 6 * 84500, beta, + 6 * 63200, gamma, + 6 * 53000, delta, + 6 * 51400, epsilon, MW of the subunits of the carbon monoxide dehydrogenase-corrinoid enzyme complex, SDS-PAGE

53000

-

6 * 19700, alpha, + 6 * 84500, beta, + 6 * 63200, gamma, + 6 * 53000, delta, + 6 * 51400, epsilon, MW of the subunits of the carbon monoxide dehydrogenase-corrinoid enzyme complex, SDS-PAGE

58000

-

x * 89000 + x * 71000 + x * 60000 + x * 58000 + x * 19000, SDS-PAGE

60000

-

x * 89000 + x * 71000 + x * 60000 + x * 58000 + x * 19000, SDS-PAGE

61600

-

gel filtration

61800

-

1 * 61800, SDS-PAGE

62000

-

2 * 62000, enzyme, + electron transfer protein CooF and membrane bound [Ni-Fe] hydrogenase, six differentsubunits; 2 * 62000, SDS-PAGE, enzyme

63200

-

6 * 19700, alpha, + 6 * 84500, beta, + 6 * 63200, gamma, + 6 * 53000, delta, + 6 * 51400, epsilon, MW of the subunits of the carbon monoxide dehydrogenase-corrinoid enzyme complex, SDS-PAGE

70000

71000

72000

-

beta subunit, SDS-PAGE

687803

73000

-

alpha2,beta2, 2 * 82000 + 2 * 73000, enzyme previously thought to have an alpha3,beta3 structure, SDS-PAGE

75000

-

L2,M2,S2, 2 * 75000 + 2 * 28400 + 2 * 17200, SDS-PAGE

78000

79400

-

2 * 19400 + 2 * 79400, SDS-PAGE

82000

-

alpha2,beta2, 2 * 82000 + 2 * 73000, enzyme previously thought to have an alpha3,beta3 structure, SDS-PAGE

84500

85970

D5G1Y2; D5G1Y0; D5G1Y1

large subunit, calculated from sequence of cDNA

712970

86600

-

1 * 86600 + 2 * 34500 + 1 * 12600, SDS-PAGE

87224

-

L2,M2,S2, 2 * 87224 + 2 * 30694 + 2 * 17752, deduced from gene sequence, SDS-PAGE

88700

-

L2,M2,S2, 2 * 17800 + 2 * 30200 + 2 * 88700, dimer of heterotrimers, SDS-PAGE, crystal structure

89000

92000

-

x * 92000 + x * 18000

94000

-

2 * 19000 + 2 * 94000, alpha2beta2, SDS-PAGE

120000

-

gel filtration, enzyme

140000

Citrobacter amalonaticus

-

gel filtration

740543

155000

-

sedimentation equilibrium centrifugation

161000

163700

-

calculated from amino acid sequence

190000

-

gel filtration

205000

-

gradient gel electrophoresis

210000

-

gel filtration

220000

-

nondenaturing PAGE

230000

-

native PAGE

232000

-

235000

-

gel filtration

250000

273500

-

deduced from amino acid sequence

390479

300000

-

sedimentation equilibrium ultracentrifugation

310000

-

685091

410000

-

gel filtration

390454, 390469

436000

-

gel filtration

450000

-

gel filtration, complex

1000000

-

the native enzyme (MW 250000 Da) forms aggregates with an MW of approximately 1000000, gradient gel electrophoresis

1600000

-

carbon monoxide dehydrogenase-corrinoid enzyme complex, gel filtration

3000000

-

high molecular weight form exists under conditions of high ionic strength, gel filtration

additional information

?

dimer

-

2 * 62000, SDS-PAGE, enzyme

heterohexamer

-

heterotetramer

hexamer

homodimer

monomer

-

1 * 61800, SDS-PAGE

octamer

-

2 * 62000, enzyme, + electron transfer protein CooF and membrane bound [Ni-Fe] hydrogenase, six differentsubunits

oligomer

-

6 * 19700, alpha, + 6 * 84500, beta, + 6 * 63200, gamma, + 6 * 53000, delta, + 6 * 51400, epsilon, MW of the subunits of the carbon monoxide dehydrogenase-corrinoid enzyme complex, SDS-PAGE

tetramer

additional information

-

properties of enzyme in complex with protein CooF which mediates electron transfer from enzyme to the CO-induced hydrogenase

672350

-

-

690057

carbon monoxide dehydrogenase II, hanging drop vapor diffusion, 20% 2-propanol, 20% polyethylene glycol 3000, 100 mM HEPES, pH 7.5

-

in presence of dithiothreitol or dithionite and under an atmosphere of N2 or CO

-

655792

-

-

native and cyanide bound CODH/ACS complex

P27989

696351

-

-

657190

vapor diffusion, 800 mM KH2PO4, 800 mM NaH2PO4, 2% 2-methyl-2,4-pentanediol, 100 mM HEPES, pH 7.3

-

EPR studies of enzyme in complex with protein CooF which mediates electron transfer from enzyme to the CO-induced hydrogenase, in presence and absence of nickel

-

672350

C56A

-

devoid of CO oxidation activity

685261

K563L

-

very low CO oxidation activity

685261

A110C

A121H/H122A

-

11% of wild type activity

A219F

-

mutant designed to block tunnel between Ni-Fe-S active site clusters, little enzymic activity. Metal clusters are properly assembled, impaired ability of CO to migrate through the tunnel

A222L

A265M

-

mutation within tunnel region of alpha subunit, absence of strong cooperative inhibition of CO, little synthesis of acetyl-CoA

A578C

-

mutant designed to block tunnel between Ni-Fe-S active site clusters, little enzymic activity. Metal clusters are properly assembled, impaired ability of CO to migrate through the tunnel

C316S

-

0.3% of wild type activity

E115H/H116A

-

24% of wild type activity

F70W

-

mutant designed to block region that connects the CO tunnel at the betabeta interface with a water channel, little enzymic activity. Metal clusters are properly assembled, impaired ability of CO to migrate through the tunnel

H113A

-

44% of wild type activity, in presence of imidazole, 45% of wild type activity

H113A/H116A/H119A

-

no activity

H113A/H119A

-

15% of wild type activity

H116A

-

6% of wild type activity, in presence of imidazole, 3% of wild type activity

H116A/G117H

-

0.8% of wild type activity

H116C

-

46% of wild type activity

H116D

-

0.4% of wild type activity

H119A

-

27% of wild type activity

H122A

-

3% of wild type activity

H122A/A123H

-

72% of wild type activity

K587A

-

42% of wild type activity

K587A/H113A

-

0.7% of wild type activity

L215F

-

mutant designed to block tunnel between Ni-Fe-S active site clusters, little enzymic activity. Metal clusters are properly assembled, impaired ability of CO to migrate through the tunnel

N101Q

-

mutant designed to block region that connects the CO tunnel at the betabeta interface with a water channel, little enzymic activity. Metal clusters are properly assembled, impaired ability of CO to migrate through the tunnel

N284A

-

41% of wild type activity

N284A/H119A

-

36% of wild type activity

C338A

-

complete loss of activity. Unable to grow in the dark with CO as the energy source, amount of enzyme present in membrane fraction is similar to wild-type as well as accumulation of Ni2+

C451A

-

complete loss of activity. Unable to grow in the dark with CO as the energy source, amount of enzyme present in membrane fraction is similar to wild-type as well as accumulation of Ni2+

C451S

-

1.4% of wild-type specific activity. Unable to grow in the dark with CO as the energy source, amount of enzyme present in membrane fraction is similar to wild-type as well as accumulation of Ni2+

C481A

-

unable to grow in the dark with CO as the energy source, amount of enzyme present in membrane fraction is similar to wild-type as well as accumulation of Ni2+

C531A

-

0.1% of wild-type specific activity

H265V

-

no CO oxidation activity in presence or absence of nickel, production of formate at same level as wild type

656296

additional information

6.5 - 9.5

-

wild-type and mutants apo-enzymes, stable for at least 30 min within this range

674921

24

-

10 min, about 30% loss of activity in presence of CO, stable in absence of CO

25 - 70

-

purification can be done at 25°C without loss of activity, heating for 30 min at 67°C leads to apparent loss, heating for only a few min at 70°C activates the enzyme

39

-

10 min, 45% loss of activity in presence of CO, stable in absence of CO

60

-

10 min, about 10% loss of activity

62

-

10 min, about 65% loss of activity in presence of CO, about 5% loss of activity in absence of CO

72

-

10 min, complete inactivation

80 - 104

-

most of the activity remains after heating at 80°C, nearly complete inactivation after heating at 104°C, greater loss of activity at all temperatures when heated in presence of CO

80

-

10 min, about 85% loss of activity in presence of CO, about 20% loss of activity in absence of CO

104

-

10 min, complete loss of activity in presence or in absence of CO

additional information

-

CO renders the enzyme more susceptible to temperature inactivation

72 h exposure to air at 4°C, about 10% loss of activity

-

98% loss of activity after 15 min in an aerobic buffer

-

CN- inactivates

-

390471

CO renders the enzyme more susceptible to temperature inactivation

-

the presence of CO has no apparent effect on the stability

-

100% of activity remains after 48 h of oxygen exposure, and after 168 h of air exposure, activity is 88% of the initial activity

-

725052

addition of the enzyme to an aerobic buffer, 50 mM Tris/HCl, pH 7.6 results in a 98% loss of activity within 15 min, presence of CO has no apparent effect on the stability of the enzyme

-

390452

enzymatic activity is destroyed by air, a low amount of activity still is present after 2 h exposure to air

-

390454

extreme oxygen lability

extremely oxygen-labile, even in N2 atmosphere with less than 5 ppm O2 the pure enzyme loses a significant amount of its activity within 18 h, 2 mM dithionite protects from inactivation

-

390450

extremely sensitive to air, most of the enzyme activity is lost upon exposure to air for 1 min, longer incubation results in complete inactivation

-

390449

oxygen stable, 10% loss in activity after 72 h exposure to air at 4°C

-

390473

Oxygen treatment results in an activity loss of 90% within 30 min. Treatment of oxygen-inactivated enzyme with CO, H2, or dithionite does not restore activity

-

727713

strong inhibition by oxygen, extremely sensitive, exposed to air activity is lost in less than 1 min

-

390449

-20°C or 5°C, 50 mM Tris/HCl, pH 7.5, 2 mM sodium dithionite, 0.2 mM methyl viologen, 50% v/v glycerol, stable for more than 1 month

-

-20°C, stored frozen in an oxygen-free atmosphere in buffer containing glycerol and 2 mM dithionite, not stable for more than 1 month

-

-70°C, samples thawed under anaerobic conditions exhibits full activity even after 6 months of storage

-

10°C, Tris/HCl, pH 7.6, enzyme is active for at least 2 days

-

4°C, 1 month, 100% inactivation

-

5°C, stored in an oxygen-free atmosphere in buffer containing glycerol and 2 mM dithionite, not stable for more than 1 month

-

-

9 entries

anion-exchange, hydrophobic interaction, gel filtration

-

carbon monoxide dehydrogenase II

-

Cosmogel His-Accept column chromatography, and gel filtration

-

724590

Ni-NTA agarose column chromatography

-

685261

Q-Sepharose column chromatography and Sephacryl S-300 gel filtration

-

Q-Sepharose column chromatography, CHT-I column chromatography, and Superdex 200 gel filtration

-

-

expressed in Escherichia coli

expressed in Escherichia coli BL21(DE3) cells

expressed in Escherichia coli DH5 alpha cells

expressed in Escherichia coli Rosetta2 (DE3) cells

-

724590

expressed in Escherichia coli strain JM109

-

685091

the synthesis of CO-DH is induced in the presence of CO. The CutR protein is involved in the induction of CO-DH synthesis in the presence of CO. When the wild type strain grown on 0.2% (w/v) glucose is further grown on 0.002% (w/v) glucose for 5 h, the CO-DH activity is increased by 1.6fold relative to that detected in the same strain grown on 0.2% (w/v) glucose for additional 5 h as a control

analysis