Table 2.

Substrate specificity of Nit09. Reactions were run for 15 min at 50 °C using 0.1 M citrate-phosphate buffer (pH 6.0), 1.25 μg purified protein, and 20 mM nitrile

Substrate	Relative activity (%)a
Phenylacetonitrile	100.00 ± 1.19
Succinonitrile	0.17 ± 0.03
Acetonitrile	0.00 ± 0.01
Crotononitrile	0.08 ± 0.01
4-Hydroxybenzonitrile	0.00 ± 0.00
Acetone cyanohydrin	0.13 ± 0.02
Cyclohexanecarbonitrile	0.09 ± 0.01
Fumaronitrile	0.45 ± 0.04
2-Phenylpropionitrile	0.52 ± 0.05
Mandelonitrile	22.01 ± 0.61
2-Phenylbutyronitrile	0.07 ± 0.01
3-Phenylpropionitrile	15.81 ± 0.80
Benzonitrile	0.48 ± 0.32
2-Naphthylacetonitrile	0.11 ± 0.00
1,4-Phenylenediacetonitrile	18.82 ± 0.62
2-Chlorophenylacetonitrile	11.69 ± 0.80
3-Chlorophenylacetonitrile	5.34 ± 0.06
4-Chlorophenylacetonitrile	4.95 ± 0.17
Cinnamonitrile	0.12 ± 0.01
3-Indoleacetonitrile	34.38 ± 0.78
2-Thiopheneacetonitrile	385.46 ± 0.78
3-Thiopheneacetonitrile	187.53 ± 5.75

^aActivity with phenylacetonitrile as substrate (5.19 U/mg) was set as 100% relative activity