Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes in Escherichia coli | Xanthobacter autotrophicus |
Protein Variants | Comment | Organism |
---|---|---|
F501H/H506E | site-directed mutagenesis of the catalytic dyad, substitution of the Phe-His active site residues by the canonical residues results in production of higher relative concentrations of acetone versus the natural product acetoacetate. Replacement of the His-Glu dyad from DSORs with Phe-His is critical for specifying carboxylation chemistry in enzyme 2-KPCC | Xanthobacter autotrophicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-(2-oxopropylthio)ethanesulfonate + CO2 + NADPH | Xanthobacter autotrophicus | - |
2-mercaptoethanesulfonate + acetoacetate + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Xanthobacter autotrophicus | Q56839 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2-mercaptoethanesulfonate + acetoacetate + NADP+ = 2-(2-oxopropylthio)ethanesulfonate + CO2 + NADPH + H+ | reaction mechanism for 2-KPCC begins with formation of enzyme-substrate disulfide adduct by the reduced form of the redox active disulfide. This step is followed by release of enolacetone anion intermediates for 2-KPCC. The primary catalytic fate of the enolacetone intermediate for the wild-type (Phe501) or variant (His501) 2-KPCC is acetoacetate or acetone, respectively | Xanthobacter autotrophicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-(2-oxopropylthio)ethanesulfonate + CO2 + NADPH | - |
Xanthobacter autotrophicus | 2-mercaptoethanesulfonate + acetoacetate + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
2-ketopropyl coenzyme M oxidoreductase/carboxylase | - |
Xanthobacter autotrophicus |
2-KPCC | - |
Xanthobacter autotrophicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Xanthobacter autotrophicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Xanthobacter autotrophicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Xanthobacter autotrophicus | |
NADPH | - |
Xanthobacter autotrophicus |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the disulfide oxidoreductase (DSOR) family of enzymes. The characteristic His-Glu catalytic dyad of the DSOR family is replaced in 2-ketopropyl coenzyme M oxidoreductase/carboxylase (2-KPCC) uniquely by the residues Phe-His, potentially to eliminate proton-donating groups at a key position in the active site. These differences in 2-KPCC are key in discriminating between carbon dioxide and protons as attacking electrophiles | Xanthobacter autotrophicus |
physiological function | 2-oxopropylcoenzyme M oxidoreductase/carboxylase (2-KPCC) catalyzes the reductive cleavage and carboxylation of 2-oxopropyl coenzyme M (2-KPC) to form acetoacetate and concomitantly regenerate CoM (2-mercaptoethanesulfonate) | Xanthobacter autotrophicus |