Crystallization (Comment) | Organism |
---|---|
native state enzyme, as well as enzyme complexed with substrate (5S)-5-amino-3-oxohexanoate or product acetoacetate, X-ray diffraction structure determination and analysis at 1.28-1.84 A resolution, molecular replacement | Candidatus Cloacimonas acidaminovorans |
Protein Variants | Comment | Organism |
---|---|---|
D231G | site-directed mutagenesis, inactive mutant | Candidatus Cloacimonas acidaminovorans |
E143G | site-directed mutagenesis | Candidatus Cloacimonas acidaminovorans |
E143Q | site-directed mutagenesis | Candidatus Cloacimonas acidaminovorans |
R226G | site-directed mutagenesis, inactive mutant | Candidatus Cloacimonas acidaminovorans |
S82G | site-directed mutagenesis | Candidatus Cloacimonas acidaminovorans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic analysis of wild-type and mutant enzymes, overview | Candidatus Cloacimonas acidaminovorans | |
0.012 | - |
acetyl-CoA | wild-type enzyme, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.012 | - |
acetyl-CoA | mutant E143Q, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.024 | - |
acetyl-CoA | mutant E143G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.046 | - |
acetyl-CoA | mutant S82G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.208 | - |
(5S)-5-amino-3-oxohexanoate | wild-type enzyme, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.442 | - |
(5S)-5-amino-3-oxohexanoate | mutant E143Q, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.921 | - |
(5S)-5-amino-3-oxohexanoate | mutant E143G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
1.181 | - |
(5S)-5-amino-3-oxohexanoate | mutant S82G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | dependent on, one metal ion per subunit and active site, coordinated by His46, His48, and Glu230 in the active site. The metal ion is situated at the bottom of a crevice accessible from two opposite openings, one of which is delimited by the mobile beta3-alpha3 loop | Candidatus Cloacimonas acidaminovorans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5S)-5-amino-3-oxohexanoate + acetyl-CoA | Candidatus Cloacimonas acidaminovorans | - |
L-3-aminobutanoyl-CoA + acetoacetate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Candidatus Cloacimonas acidaminovorans | B0VHH0 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(5S)-5-amino-3-oxohexanoate + acetyl-CoA = L-3-aminobutanoyl-CoA + acetoacetate | the enzyme shows a catalytic reaction mechanism that proceeds through deprotonation of the 3-keto-5-aminohexanoate substrate, nucleophilic addition onto an incoming acetyl-CoA, intramolecular transfer of the CoA moiety, and final retro-Claisen reaction leading to acetoacetate and 3-aminobutyryl-CoA, structure-function analysis, docking study and molecular modeling, detailed overview. Absence of detection of any covalent acyl-enzyme intermediate | Candidatus Cloacimonas acidaminovorans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5S)-5-amino-3-oxohexanoate + acetyl-CoA | - |
Candidatus Cloacimonas acidaminovorans | L-3-aminobutanoyl-CoA + acetoacetate | - |
r |
Subunits | Comment | Organism |
---|---|---|
homotetramer | each subunit of 276 residues shows the canonical (beta/alpha)8 TIM barrel fold, with a short additional C-terminal alpha-helix (alpa9) protruding at the N-terminal face of the barrel and three beta-turn extensions coming out from the barrel core, inserted in between beta2 and alpha2 (residues 49-58), beta4 and alpha4 (residues 112-119), and beta8 and alpha8 (residues 234-240) | Candidatus Cloacimonas acidaminovorans |
Synonyms | Comment | Organism |
---|---|---|
kce | - |
Candidatus Cloacimonas acidaminovorans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.13 | - |
acetyl-CoA | mutant S82G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.13 | - |
(5S)-5-amino-3-oxohexanoate | mutant S82G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.32 | - |
acetyl-CoA | mutant E143Q, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.32 | - |
(5S)-5-amino-3-oxohexanoate | mutant E143Q, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.37 | - |
acetyl-CoA | mutant E143G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.37 | - |
(5S)-5-amino-3-oxohexanoate | mutant E143G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
2.69 | - |
acetyl-CoA | wild-type enzyme, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
2.69 | - |
(5S)-5-amino-3-oxohexanoate | wild-type enzyme, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a representative of a large family of prokaryotic hypothetical proteins, annotated as the domain of unknown function DUF849, it shows the ubiquitous triose phosphate isomerase (TIM) barrel fold and a Zn2+ cation reminiscent of metal-dependent class II aldolases | Candidatus Cloacimonas acidaminovorans |
metabolism | the enzyme is involved in the anaerobic fermentation of lysine | Candidatus Cloacimonas acidaminovorans |
additional information | enzyme structure determination and analysis, the active site is situated in each monomer at the C-terminal face of the central beta-barrel with a metal ion coordinated by His46, His48, and Glu230, overview | Candidatus Cloacimonas acidaminovorans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
(5S)-5-amino-3-oxohexanoate | mutant S82G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.4 | - |
(5S)-5-amino-3-oxohexanoate | mutant E143G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
0.7 | - |
(5S)-5-amino-3-oxohexanoate | mutant E143Q, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
2.8 | - |
acetyl-CoA | mutant S82G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
12.9 | - |
(5S)-5-amino-3-oxohexanoate | wild-type enzyme, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
15.4 | - |
acetyl-CoA | mutant E143G, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
26.7 | - |
acetyl-CoA | mutant E143Q, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans | |
224.2 | - |
acetyl-CoA | wild-type enzyme, pH and temperature not specified in the publication | Candidatus Cloacimonas acidaminovorans |