Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli | Thermobifida fusca |
Protein Variants | Comment | Organism |
---|---|---|
additional information | addition of the dimerization-docking and anchoring domain (RIDD-RIAD) system to the C-terminus of the NADPH-dependent Baeyer-Villiger monooxygenase phenylacetone monooxygenase (PAMO) and the NADPH-regenerating enzyme (phosphite dehydrogenase, PTDH, EC 1.20.1.1) allowing self-assembly based on specific protein-protein interactions between both peptides and allow tuning of the ratio of the targeted enzymes as the RIAD peptide binds to two RIDD peptides. Several RIDD/RIAD-tagged PAMO and PTDH variants are successfully overproduced in Escherichia coli and subsequently purified. Complementary tagged enzymes are mixed and analyzed for their oligomeric state, stability, and activity. Complexes are formed in the case of some specific combinations (PAMORIAD-PTDHRIDD and PAMORIAD/RIAD-PTDHRIDD). These enzyme complexes display similar catalytic activity when compared with the PTDH-PAMO fusion enzyme. The thermostability of PAMO in these complexes is retained while PTDH displays somewhat lower thermostability | Thermobifida fusca |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
phenylacetone + NADPH + H+ + O2 | Thermobifida fusca | - |
benzyl acetate + NADP+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermobifida fusca | Q47PU3 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
phenylacetone + NADPH + H+ + O2 | - |
Thermobifida fusca | benzyl acetate + NADP+ + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PAMO | - |
Thermobifida fusca |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Thermobifida fusca |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Thermobifida fusca |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Thermobifida fusca | |
NADPH | - |
Thermobifida fusca |
General Information | Comment | Organism |
---|---|---|
evolution | phenylacetone monooxygenase, PAMO, is a NADPH-dependent Baeyer-Villiger monooxygenase | Thermobifida fusca |