Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | enzyme activation mechanism, overview | Cupriavidus necator | |
NADH | specific activation of the soluble oligomeric enzyme | Cupriavidus necator | |
NADH | activates hexameric enzyme form, no activation of the tetrameric enzyme form | Cupriavidus necator | |
NADPH | specific activation of the soluble oligomeric enzyme, the binding domain is located on subunit HoxI | Cupriavidus necator | |
NADPH | activates hexameric enzyme form. Subunit HoxI provides a binding domain for NADPH | Cupriavidus necator |
Cloned (Comment) | Organism |
---|---|
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 |
Protein Variants | Comment | Organism |
---|---|---|
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 |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytoplasm | - |
Cupriavidus necator | 5737 | - |
soluble | - |
Cupriavidus necator | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
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 | |
CO | the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre | Cupriavidus necator | |
Fe2+ | the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre | Cupriavidus necator | |
Ni2+ | the active site is a (enzyme-Cys)2(CN)Ni(micro-enzyme-Cys)2Fe(CN)3(CO) centre | Cupriavidus necator |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
H2 + NAD+ | Cupriavidus necator | key enzyme in H2 metabolism | H+ + NADH | - |
r | |
H2 + NAD+ | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | key enzyme in H2 metabolism | H+ + NADH | - |
r | |
additional information | Cupriavidus necator | the organism can grow on H2 as sole energy source in an oxic environment | ? | - |
? | |
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 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cupriavidus necator | - |
- |
- |
Cupriavidus necator | - |
facultative lithoautotrophic proteobacterium, formerly Alcaligenes eutrophus | - |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | - |
- |
- |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | - |
facultative lithoautotrophic proteobacterium, formerly Alcaligenes eutrophus | - |
Purification (Comment) | Organism |
---|---|
hexameric enzyme form | Cupriavidus necator |
recombinant Strep-tagged HoxI-deletion mutant tetramer from Escherichia coli by 2 different procedures | Cupriavidus necator |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Cupriavidus necator |
0.6 | - |
reaction with NADPH and ferrocyanide, purified wild-type enzyme complex including HoxI, no reaction with the HoxI deletion mutant | Cupriavidus necator |
25.2 | - |
reaction with H2 and benzyl viologen, purified HoxI deletion mutant enzyme | Cupriavidus necator |
35.1 | - |
reaction with H2 and benzyl viologen, purified wild-type enzyme complex including HoxI | Cupriavidus necator |
39 | - |
reaction with H2 and NAD+, purified HoxI deletion mutant enzyme | Cupriavidus necator |
41.9 | - |
reaction with H2 and NAD+, purified wild-type enzyme complex including HoxI | Cupriavidus necator |
61.6 | - |
reaction with NADH and ferrocyanide, purified HoxI deletion mutant enzyme | Cupriavidus necator |
64.9 | - |
reaction with NADH and ferrocyanide, purified wild-type enzyme complex including HoxI | Cupriavidus necator |
102.1 | - |
reaction with H2 and ferrocyanide, purified HoxI deletion mutant enzyme | Cupriavidus necator |
108.8 | - |
reaction with H2 and ferrocyanide, purified wild-type enzyme complex including HoxI | Cupriavidus necator |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ferrocyanide + NAD(P)H | enzyme complex including HoxI | Cupriavidus necator | ferricyanide + NAD(P)+ | - |
? | |
H2 + ferrocyanide | - |
Cupriavidus necator | H+ + ferricyanide | - |
r | |
H2 + NAD+ | - |
Cupriavidus necator | H+ + NADH | - |
r | |
H2 + NAD+ | no activity with NADP+ | Cupriavidus necator | H+ + NADH | - |
r | |
H2 + NAD+ | key enzyme in H2 metabolism | Cupriavidus necator | H+ + NADH | - |
r | |
H2 + NAD+ | - |
Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | H+ + NADH | - |
r | |
H2 + NAD+ | no activity with NADP+ | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | H+ + NADH | - |
r | |
H2 + NAD+ | key enzyme in H2 metabolism | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | H+ + NADH | - |
r | |
H2 + oxidized benzyl viologen | - |
Cupriavidus necator | H+ + reduced benzyl viologen | - |
r | |
H2 + oxidized benzyl viologen | - |
Cupriavidus necator | reduced benzyl viologen + H+ | - |
? | |
additional information | the organism can grow on H2 as sole energy source in an oxic environment | Cupriavidus necator | ? | - |
? | |
additional information | tetrameric or hexameric enzyme form: no H2 production from NADPH | Cupriavidus necator | ? | - |
? | |
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 | ? | - |
? | |
additional information | tetrameric or hexameric enzyme form: no H2 production from NADPH | Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 | ? | - |
? | |
NADH + K3Fe(CN)6 | - |
Cupriavidus necator | ? | - |
? | |
NADPH + K3Fe(CN)6 | low reaction with hexameric enzyme form, no reaction with tetrameric enzyme form | Cupriavidus necator | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | subunit stoichiometry of HoxFUYI2. Subunit HoxIhas a MW of 19000 Da as determined by SDS-PAGE | Cupriavidus necator |
More | structure analysis and subunit composition, HoxI is associated with the NADH-dehydrogenase moiety of the enzyme | Cupriavidus necator |
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 |
Synonyms | Comment | Organism |
---|---|---|
SH | - |
Cupriavidus necator |
soluble [NiFe]-hydrogenase | - |
Cupriavidus necator |
[NiFe]-hydrogenase | - |
Cupriavidus necator |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | - |
NADPH | pH 8.0, wild-type enzyme complex including HoxI, with ferrocyanide | Cupriavidus necator | |
2 | - |
ferrocyanide | pH 8.0, wild-type enzyme complex including HoxI, with NADPH | Cupriavidus necator | |
70 | - |
Oxidized benzyl viologen | pH 8.0, HoxI deletion mutant enzyme, with H2 | Cupriavidus necator | |
70 | - |
H2 | pH 8.0, HoxI deletion mutant enzyme, with oxidized benzyl viologen | Cupriavidus necator | |
109 | - |
NAD+ | pH 8.0, HoxI deletion mutant enzyme, with H2 | Cupriavidus necator | |
109 | - |
H2 | pH 8.0, HoxI deletion mutant enzyme, with NAD+ | Cupriavidus necator | |
120 | - |
Oxidized benzyl viologen | pH 8.0, wild-type enzyme complex including HoxI, with H2 | Cupriavidus necator | |
120 | - |
H2 | pH 8.0, wild-type enzyme complex including HoxI, with oxidized benzyl viologen | Cupriavidus necator | |
143 | - |
NAD+ | pH 8.0, wild-type enzyme complex including HoxI, with H2 | Cupriavidus necator | |
143 | - |
H2 | pH 8.0, wild-type enzyme complex including HoxI, with NAD+ | Cupriavidus necator | |
171 | - |
NADH | pH 8.0, HoxI deletion mutant enzyme, with ferrocyanide | Cupriavidus necator | |
171 | - |
ferrocyanide | pH 8.0, HoxI deletion mutant enzyme, with NADH | Cupriavidus necator | |
171 | - |
NADH | reaction with K3Fe(CN)6, tetrameric enzyme form | Cupriavidus necator | |
222 | - |
NADH | pH 8.0, wild-type enzyme complex including HoxI, with ferrocyanide | Cupriavidus necator | |
222 | - |
ferrocyanide | pH 8.0, wild-type enzyme complex including HoxI, with NADH | Cupriavidus necator | |
222 | - |
NADH | reaction with K3Fe(CN)6, hexameric enzyme form | Cupriavidus necator | |
284 | - |
H2 | pH 8.0, HoxI deletion mutant enzyme, with ferrocyanide | Cupriavidus necator | |
284 | - |
ferrocyanide | pH 8.0, HoxI deletion mutant enzyme, with H2 | Cupriavidus necator | |
372 | - |
H2 | pH 8.0, wild-type enzyme complex including HoxI, with ferrocyanide | Cupriavidus necator | |
372 | - |
ferrocyanide | pH 8.0, wild-type enzyme complex including HoxI, with H2 | Cupriavidus necator |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Cupriavidus necator |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
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 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | no activity with NADP+ | Cupriavidus necator | |
NAD+ | - |
Cupriavidus necator |