3.5.1.46	A61V/A124V/R187S/F264C/G291R/G338A/D370Y	site-directed mutagenesis	753567	
3.5.1.46	A61V/A253T/F264C/D370Y	site-directed mutagenesis, 10fold increased activity with N-(6-aminohexanoyl)-6-aminohexanoate	668639	
3.5.1.46	D181E	mutant with increased Km and decreased activity	-, 172041	
3.5.1.46	D181E	site-directed mutagenesis of EII, the mutant shows reduced activity compared to the wild-type enzyme	669406	
3.5.1.46	D181H	-	-, 172041	
3.5.1.46	D181H	site-directed mutagenesis of EII, the mutant shows highly reduced activity compared to the wild-type enzyme	669406	
3.5.1.46	D181K	-	-, 172041	
3.5.1.46	D181K	site-directed mutagenesis of EII, nearly inactive mutant	669406	
3.5.1.46	D181N	mutant with increased Km and decreased activity	-, 172041	
3.5.1.46	D181N	site-directed mutagenesis of EII, the mutant shows reduced activity compared to the wild-type enzyme	669406	
3.5.1.46	G181D/H266N	construction of NylB mutant enzyme Hyb-24DN, the activity in Hyb-24 is enhanced 150fold by the two substitutions, where Asp181-COO- stabilizes the substrate binding by electrostatic interactions with Ald-NH3+, and Asn266 cooperatively improves the electrostatic environment with Asp181	753567	
3.5.1.46	additional information	construction of chimeric NylB/NylB' mutants Hyb-24DN and Hyb-24DNY. A NylB/NylB' hybrid enzyme Hyb-24 (NylB' containing T3A-P4R-T5S-S8Q-D15G substitutions) has about 0.5% of the NylB level of Ald-hydrolytic activity. Surface structure of the entrance of the catalytic cleft of the Hyb-24DNY and its mutant enzymes, wild-type and mutant kinetics, detailed overview	753567	
3.5.1.46	R187A	site-directed mutagenesis, the substitution in the parental Hyb-24DNY decreases the specific activity (initial reaction rate) of synthesis to 37% of the level of Hyb-24DNY	753567	
3.5.1.46	R187G	site-directed mutagenesis, the substitution in the parental Hyb-24DNY decreases the specific activity (initial reaction rate) of synthesis to 15% of the level of Hyb-24DNY	753567	
3.5.1.46	R187S	site-directed mutagenesis, the substitution in the parental Hyb-24DNY decreases the specific activity (initial reaction rate) of synthesis to 24% of the level of Hyb-24DNY	753567	
3.5.1.46	R187S/F264C/D370Y	site-directed mutagenesis, the substitutions also enhance the Ald-hydrolytic activity 80fold over the parental Hyb-24 enzyme. The directly evolved enzyme (Hyb-S4M94) possesses almost no Ald-synthetic activity, although it possesses high levels of the hydrolytic activity even in 90% t-butyl alcohol. In this mutant enzyme, the R187S/F264C substitutions stabilizes the binding at the N-terminal region of Ald, while D370Y substitution stabilizes the binding at the C-terminal region of Ald	753567	
3.5.1.46	T3A/P4R/T5S/S8Q/D15G	construction of a hybrid of isozymes EII and EII', termed Hyb24, by five amino acid replacement in EII', the mutant shows the same activity as EII'	667074, 669406	
3.5.1.46	T3A/P4R/T5S/S8Q/D15G	site-directed mutagenesis, construction of a hybrid of isozymes EII and EII', termed Hyb24, by five amino acid replacement in EII', the mutant shows the same activity as EII'	668639	
3.5.1.46	T3A/P4R/T5S/S8Q/D15G/D370Y	site-directed mutagenesis, mutant Hyb24, by five amino acid replacement in EII' for residues of EII, plus 2 additional exchanges for EII residues leading to a 10fold increased activity with N-(6-aminohexanoyl)-6-aminohexanoate	668639	
3.5.1.46	T3A/P4R/T5S/S8Q/D15G/G181D	site-directed mutagenesis, mutant Hyb24, by five amino acid replacement in EII' for residues of EII, plus 2 additional exchanges for EII residues leading to a 10fold increased activity with N-(6-aminohexanoyl)-6-aminohexanoate	668639	
3.5.1.46	T3A/P4R/T5S/S8Q/D15G/G181D/D370Y	site-directed mutagenesis, mutant Hyb24, by five amino acid replacement in EII' for residues of EII, plus 2 additional exchanges for EII residues leading to a 100fold increased activity with N-(6-aminohexanoyl)-6-aminohexanoate	668639