EC Number |
Protein Variants |
Reference |
---|
1.8.1.5 | C82A |
mutagenesis of the interchange thiol, abolishes all redox-dependent reactions. Redox-independent acetoacetate decarboxylation is not decreased |
725265 |
1.8.1.5 | C87A |
mutagenesis of the flavin thiol, results in an inactive enzyme for steady-state redox-dependent reactions, but this variant catalyzes a single-turnover reaction producing a 0.8:1 ratio of product to enzyme. Redox-independent acetoacetate decarboxylation is not decreased |
725265 |
1.8.1.5 | F501H |
the mutant shows 10% acetoacetate production activity compared to the wild type enzyme. The overall rate of NADPH turnover remains relatively unchanged in the F501H variant relative to wild type. Moreover, acetone formation by F501H is comparable in rate to the carboxylation reaction catalyzed by wild type enzyme and leading to acetoacetate |
765078 |
1.8.1.5 | F501H/H506E |
inactive |
765078 |
1.8.1.5 | 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 |
742523 |
1.8.1.5 | H137A |
mutagenesis of the histidine proximal to the ordered water molecule, leads to nearly complete loss of redox-dependent reactions. Redox-independent acetoacetate decarboxylation is not decreased |
725265 |
1.8.1.5 | H506E |
the mutant shows 37% acetoacetate production activity compared to the wild type enzyme. NADPH turnover is around 1.5fold slower in H506E versus wild type enzyme |
765078 |
1.8.1.5 | H84A |
mutagenesis of the distal histidine residue, reduces the redox-dependent activities by 58 to 76%. Redox-independent acetoacetate decarboxylation is not decreased |
725265 |
1.8.1.5 | M140A |
residue flanking the substrate, catalytic efficiency for 2-(2-oxopropylthio)ethanesulfonate carboxylation is 47fold lower than that for wild-type |
725265 |