EC Number | Activating Compound | Comment | Organism | Structure |
---|---|---|---|---|
1.12.1.2 | additional information | enzyme activation mechanism, overview | Cupriavidus necator | |
1.12.1.2 | NADH | specific activation of the soluble oligomeric enzyme | Cupriavidus necator | |
1.12.1.2 | NADH | activates hexameric enzyme form, no activation of the tetrameric enzyme form | Cupriavidus necator | |
1.12.1.2 | NADPH | specific activation of the soluble oligomeric enzyme, the binding domain is located on subunit HoxI | Cupriavidus necator | |
1.12.1.2 | NADPH | activates hexameric enzyme form. Subunit HoxI provides a binding domain for NADPH | Cupriavidus necator |
EC Number | Cloned (Comment) | Organism |
---|---|---|
1.12.1.2 | gene cluster HoxFUYHI, cloning and expression of the Strep-tagged HoxI-deletion mutant, a tetramer composed of HoxFU and HoxHY dimers forming a tetramer in Escherichia coli strains JM109 and XL-1 Blue | Cupriavidus necator |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.12.1.2 | additional information | construction of a HoxI deletion mutant enzyme, which shows different activation behaviour than the wild-type, it is not activated by and does not react with NADPH | Cupriavidus necator |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
1.12.1.2 | cytoplasm | - |
Cupriavidus necator | 5737 | - |
1.12.1.2 | soluble | - |
Cupriavidus necator | - |
- |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.12.1.2 | CN- | enzyme contains four cyanides in its active site, one is bound to the Ni2+, the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre, the CN- bound to the nickel ion can be irreversibly removed inducing enzyme inhibition by oxygen | Cupriavidus necator | |
1.12.1.2 | CO | the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre | Cupriavidus necator | |
1.12.1.2 | Fe2+ | the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre | Cupriavidus necator | |
1.12.1.2 | Ni2+ | the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre | Cupriavidus necator |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.12.1.2 | H2 + NAD+ | Cupriavidus necator | key enzyme in H2 metabolism | H+ + NADH | - |
r | |
1.12.1.2 | H2 + NAD+ | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | key enzyme in H2 metabolism | H+ + NADH | - |
r | |
1.12.1.2 | additional information | Cupriavidus necator | the organism can grow on H2 as sole energy source in an oxic environment | ? | - |
? | |
1.12.1.2 | additional information | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | the organism can grow on H2 as sole energy source in an oxic environment | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.12.1.2 | Cupriavidus necator | - |
- |
- |
1.12.1.2 | Cupriavidus necator | - |
facultative lithoautotrophic proteobacterium, formerly Alcaligenes eutrophus | - |
1.12.1.2 | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | - |
- |
- |
1.12.1.2 | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | - |
facultative lithoautotrophic proteobacterium, formerly Alcaligenes eutrophus | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.12.1.2 | hexameric enzyme form | Cupriavidus necator |
1.12.1.2 | recombinant Strep-tagged HoxI-deletion mutant tetramer from Escherichia coli by 2 different procedures | Cupriavidus necator |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
1.12.1.2 | additional information | - |
- |
Cupriavidus necator |
1.12.1.2 | 0.6 | - |
reaction with NADPH and ferrocyanide, purified wild-type enzyme complex including HoxI, no reaction with the HoxI deletion mutant | Cupriavidus necator |
1.12.1.2 | 25.2 | - |
reaction with H2 and benzyl viologen, purified HoxI deletion mutant enzyme | Cupriavidus necator |
1.12.1.2 | 35.1 | - |
reaction with H2 and benzyl viologen, purified wild-type enzyme complex including HoxI | Cupriavidus necator |
1.12.1.2 | 39 | - |
reaction with H2 and NAD+, purified HoxI deletion mutant enzyme | Cupriavidus necator |
1.12.1.2 | 41.9 | - |
reaction with H2 and NAD+, purified wild-type enzyme complex including HoxI | Cupriavidus necator |
1.12.1.2 | 61.6 | - |
reaction with NADH and ferrocyanide, purified HoxI deletion mutant enzyme | Cupriavidus necator |
1.12.1.2 | 64.9 | - |
reaction with NADH and ferrocyanide, purified wild-type enzyme complex including HoxI | Cupriavidus necator |
1.12.1.2 | 102.1 | - |
reaction with H2 and ferrocyanide, purified HoxI deletion mutant enzyme | Cupriavidus necator |
1.12.1.2 | 108.8 | - |
reaction with H2 and ferrocyanide, purified wild-type enzyme complex including HoxI | Cupriavidus necator |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.12.1.2 | ferrocyanide + NAD(P)H | enzyme complex including HoxI | Cupriavidus necator | ferricyanide + NAD(P)+ | - |
? | |
1.12.1.2 | H2 + ferrocyanide | - |
Cupriavidus necator | H+ + ferricyanide | - |
r | |
1.12.1.2 | H2 + NAD+ | - |
Cupriavidus necator | H+ + NADH | - |
r | |
1.12.1.2 | H2 + NAD+ | no activity with NADP+ | Cupriavidus necator | H+ + NADH | - |
r | |
1.12.1.2 | H2 + NAD+ | key enzyme in H2 metabolism | Cupriavidus necator | H+ + NADH | - |
r | |
1.12.1.2 | H2 + NAD+ | - |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | H+ + NADH | - |
r | |
1.12.1.2 | H2 + NAD+ | no activity with NADP+ | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | H+ + NADH | - |
r | |
1.12.1.2 | H2 + NAD+ | key enzyme in H2 metabolism | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | H+ + NADH | - |
r | |
1.12.1.2 | H2 + oxidized benzyl viologen | - |
Cupriavidus necator | H+ + reduced benzyl viologen | - |
r | |
1.12.1.2 | H2 + oxidized benzyl viologen | - |
Cupriavidus necator | reduced benzyl viologen + H+ | - |
? | |
1.12.1.2 | additional information | the organism can grow on H2 as sole energy source in an oxic environment | Cupriavidus necator | ? | - |
? | |
1.12.1.2 | additional information | tetrameric or hexameric enzyme form: no H2 production from NADPH | Cupriavidus necator | ? | - |
? | |
1.12.1.2 | additional information | the organism can grow on H2 as sole energy source in an oxic environment | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | ? | - |
? | |
1.12.1.2 | additional information | tetrameric or hexameric enzyme form: no H2 production from NADPH | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | ? | - |
? | |
1.12.1.2 | NADH + K3Fe(CN)6 | - |
Cupriavidus necator | ? | - |
? | |
1.12.1.2 | NADPH + K3Fe(CN)6 | low reaction with hexameric enzyme form, no reaction with tetrameric enzyme form | Cupriavidus necator | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.12.1.2 | hexamer | subunit stoichiometry of HoxFUYI2. Subunit HoxIhas a MW of 19000 Da as determined by SDS-PAGE | Cupriavidus necator |
1.12.1.2 | More | structure analysis and subunit composition, HoxI is associated with the NADH-dehydrogenase moiety of the enzyme | Cupriavidus necator |
1.12.1.2 | oligomer | enzyme is composed as a dimer of pentamers, the latter consisting of subunit pairs HoxU with HoxF, and HoxY with HoxH, and a fifth 19 kDa subunit HoxI, i.e. B protein, the HoxFU dimer is the NADH-dehydrogenase moiety, the HoxHY dimer is the hydrogenase moiety, overview | Cupriavidus necator |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.12.1.2 | SH | - |
Cupriavidus necator |
1.12.1.2 | soluble [NiFe]-hydrogenase | - |
Cupriavidus necator |
1.12.1.2 | [NiFe]-hydrogenase | - |
Cupriavidus necator |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.12.1.2 | 2 | - |
NADPH | pH 8.0, wild-type enzyme complex including HoxI, with ferrocyanide | Cupriavidus necator | |
1.12.1.2 | 2 | - |
ferrocyanide | pH 8.0, wild-type enzyme complex including HoxI, with NADPH | Cupriavidus necator | |
1.12.1.2 | 70 | - |
Oxidized benzyl viologen | pH 8.0, HoxI deletion mutant enzyme, with H2 | Cupriavidus necator | |
1.12.1.2 | 70 | - |
H2 | pH 8.0, HoxI deletion mutant enzyme, with oxidized benzyl viologen | Cupriavidus necator | |
1.12.1.2 | 109 | - |
NAD+ | pH 8.0, HoxI deletion mutant enzyme, with H2 | Cupriavidus necator | |
1.12.1.2 | 109 | - |
H2 | pH 8.0, HoxI deletion mutant enzyme, with NAD+ | Cupriavidus necator | |
1.12.1.2 | 120 | - |
Oxidized benzyl viologen | pH 8.0, wild-type enzyme complex including HoxI, with H2 | Cupriavidus necator | |
1.12.1.2 | 120 | - |
H2 | pH 8.0, wild-type enzyme complex including HoxI, with oxidized benzyl viologen | Cupriavidus necator | |
1.12.1.2 | 143 | - |
NAD+ | pH 8.0, wild-type enzyme complex including HoxI, with H2 | Cupriavidus necator | |
1.12.1.2 | 143 | - |
H2 | pH 8.0, wild-type enzyme complex including HoxI, with NAD+ | Cupriavidus necator | |
1.12.1.2 | 171 | - |
NADH | pH 8.0, HoxI deletion mutant enzyme, with ferrocyanide | Cupriavidus necator | |
1.12.1.2 | 171 | - |
ferrocyanide | pH 8.0, HoxI deletion mutant enzyme, with NADH | Cupriavidus necator | |
1.12.1.2 | 171 | - |
NADH | reaction with K3Fe(CN)6, tetrameric enzyme form | Cupriavidus necator | |
1.12.1.2 | 222 | - |
NADH | pH 8.0, wild-type enzyme complex including HoxI, with ferrocyanide | Cupriavidus necator | |
1.12.1.2 | 222 | - |
ferrocyanide | pH 8.0, wild-type enzyme complex including HoxI, with NADH | Cupriavidus necator | |
1.12.1.2 | 222 | - |
NADH | reaction with K3Fe(CN)6, hexameric enzyme form | Cupriavidus necator | |
1.12.1.2 | 284 | - |
H2 | pH 8.0, HoxI deletion mutant enzyme, with ferrocyanide | Cupriavidus necator | |
1.12.1.2 | 284 | - |
ferrocyanide | pH 8.0, HoxI deletion mutant enzyme, with H2 | Cupriavidus necator | |
1.12.1.2 | 372 | - |
H2 | pH 8.0, wild-type enzyme complex including HoxI, with ferrocyanide | Cupriavidus necator | |
1.12.1.2 | 372 | - |
ferrocyanide | pH 8.0, wild-type enzyme complex including HoxI, with H2 | Cupriavidus necator |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.12.1.2 | 8 | - |
assay at | Cupriavidus necator |
EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|---|
1.12.1.2 | 6.2 | 7 | stability of the enzyme complex including HoxI in potassium phosphate buffer at pH 6.2 and pH 7.0, not at pH 8.0 or in Tris-HCl buffer at pH 7.2 and pH 8.0, or in Tris-nitrate buffer at pH 7.4 | Cupriavidus necator |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.12.1.2 | additional information | no activity with NADP+ | Cupriavidus necator | |
1.12.1.2 | NAD+ | - |
Cupriavidus necator |