EC Number |
Protein Variants |
Reference |
---|
1.1.1.B4 | A90S |
activity with NADPH as cofactor decreased to about 50% of wild-type, activity with NADH strongly decreased |
711860 |
1.1.1.B4 | C295A |
the mutant shows an increased size of the alkyl group which can bind in the substrate small pocket by one carbon atom compared to the wild-type enzyme |
741784 |
1.1.1.B4 | G37D/R38P |
activity with NADH is decreased relative to Arg38Pro alone, but is higher than that of the wild-type enzyme |
711860 |
1.1.1.B4 | I86A |
site-directed mutagensis, the secondary alcohol dehydrogenase I86A mutant is stereospecific for (R)-alcohols instead of (S)-alcohols, in contrast to the wild-type enzyme, the mutation I86A allows large substituents to fit into the large pocket of I86ATeSADH, which corresponds to the small pocket in wild-type TeSADH, modeling of the stereopreference of TeSADH I86A |
723841 |
1.1.1.B4 | I86A |
the mutant shows altered substrate specificity and expands substrate specificity to include acetophenone, which is a very poor substrate for wild-type SADH, but also reverses the usual preferred stereochemistry to produce the anti-prelog R-product. I86A SADH exhibits limited reactivity with substituted acetophenones, fluorine being the only tolerable substituent |
741784 |
1.1.1.B4 | I86A/C295A |
the mutant enzyme shows broadened substrate specificity for aryl ketones and broadened substrate specificity for meta-substituted, but not para-substituted, acetophenones compared to the wild-type enzyme. The increase of the substrate specificity of I86A/C295A SADH is accompanied by a decrease in the kcat/Km values of acetophenones, possibly due to the substrates fitting loosely inside the more open active site |
741784 |
1.1.1.B4 | M140I |
no activity with NADPH as cofactor |
711860 |
1.1.1.B4 | R38P |
no activity with NADPH as cofactor, fourfold increase in activity with NADH |
711860 |
1.1.1.B4 | S154Y |
mutant exhibits nearly 13fold, 5.4fold, and 2.3fold increase in kcat/Km value, kcat value, and specific activity toward 3,5-bis(trifluoromethyl)acetophenone |
-, 739942 |
1.1.1.B4 | S154Y/L194I |
180% of wild-type activity |
-, 739942 |