EC Number   |
Protein Variants |
Reference |
|---|
   1.14.13.92 | more |
a rational approach is used to improve the robustness of enzymes, in particular, Baeyer-Villiger monooxygenases (BVMOs) against H2O2. The enzyme access tunnels, which may serve as exit paths for H2O2 from the active site to the bulk, are predicted by using the CAVER and/or protein energy landscape exploration (PELE) software for mutant PAMO_C65D from Thermobifida fusca. The amino acid residues, which are susceptible to oxidation by H2O2 (e.g. methionine and tyrosine) and located in vicinity of the predicted H2O2 migration paths, are substituted with less reactive or inert amino acids (e.g. leucine and isoleucine), leading to design of H2O2-resistant enzyme variants |
763862 |
| 1.14.13.92 | more |
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 |
764264 |
| 1.14.13.92 | more |
construction of three mutants P1-P3 by elimination of a bulge loop region, involving residues Ser441, Ala442, and Leu443, leading to enhanced substrate enantioselectivity of Baeyer-Villiger reactions while maintaining high thermal stability, overview |
671189 |
| 1.14.13.92 | more |
directed evolution of phenylacetone monooxygenase as an active catalyst for the Baeyer-Villiger conversion of cyclohexanone to caprolactone using iterative saturation mutagenesis, mutant screening, overview. Molecular dynamics simulations and induced fit docking of wild-type and mutant enzymes with cyclohexanone. The mutants are used in the whole cell system of Escherichia coli cells |
744548 |
| 1.14.13.92 | more |
engineering of three highly stereoselective mutants of the thermally stable phenylacetone monooxygenase as practical catalysts for enantioselective Baeyer-Villiger oxidations of several ketones on a preparative scale under in vitro conditions, optimization of the method including a coupled cofactor-regeneration system, reaction mechanism, overview |
671682 |
| 1.14.13.92 | M446G |
enzyme has altered activity and substrate specificity |
697754 |
| 1.14.13.92 | more |
expanding the substrate scope of a thermostable phenylacetone monooxygenase (PAMO) to cyclohexanone by using site-directed mutagenesis. Several mutants are found to be active with cyclohexanone for which wild-type PAMO shows no activity. There is possible additive or cooperative effect existing between I67 and P440. Based on the thermostable PAMO scaffold, a chimeric PAMO-CHMO enzyme mutant is created, which shows no activity on cyclohexanone |
764409 |
| 1.14.13.92 | P440F |
higher subtrate variability, temperature optimum at 50C with range from 45-56C |
698487 |
| 1.14.13.92 | P440H |
higher subtrate variability, temperature optimum at 50C with range from 45-56C |
698487 |
| 1.14.13.92 | P440I |
higher subtrate variability, temperature optimum at 50C with range from 45-56C |
698487 |
