Literature summary for 4.2.1.84 extracted from

Song, L.; Wang, M.; Yang, X.; Qian, S.
Purification and characterization of the enantioselective nitrile hydratase from Rhodococcus sp. AJ270 (2007), Biotechnol. J., 2, 717-724.

Inhibitors

Inhibitors	Comment	Organism
AgNO3	1 mM, complete inhibition of the enzyme	Rhodococcus sp.
ammonium persulfate	1 mM, relative activity: 0.3%	Rhodococcus sp.
CuCl2	1 mM, relative activity: 4.8%	Rhodococcus sp.
EDTA	1 mM, relative activity: 96.2%	Rhodococcus sp.
hydrogen peroxide	1 mM, relative activity: 0%	Rhodococcus sp.
iodoacetamide	1 mM, relative activity: 68.6%	Rhodococcus sp.
iodoacetic acid	1 mM, relative activity: 93.0%	Rhodococcus sp.
LiCl	1 mM, relative activity: 97.3%	Rhodococcus sp.
MgCl2	1 mM, relative activity: 73.7%	Rhodococcus sp.
N-bromosuccinimide	1 mM, relative activity: 0.2%	Rhodococcus sp.
Sodium azide	1 mM, relative activity: 61.3%	Rhodococcus sp.
ZnCl2	1 mM, relative activity: 77.6%	Rhodococcus sp.
ZnSO4	1 mM, relative activity: 99.5%	Rhodococcus sp.

Metals/Ions

Metals/Ions	Comment	Organism
BaCl2	1 mM, relative activity: 101.1%	Rhodococcus sp.
CaCl2	1 mM, relative activity: 102.0%	Rhodococcus sp.
CoCl2	1 mM, relative activity: 106.4%	Rhodococcus sp.
Na2MoO4	1 mM, relative activity: 102.1%	Rhodococcus sp.

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
22975	-	alphabeta, 1 * 22975 + 1 * 23493, MALDI-TOF MS	Rhodococcus sp.
23493	-	alphabeta, 1 * 22975 + 1 * 23493, MALDI-TOF MS	Rhodococcus sp.

Organism

Organism	UniProt	Comment	Textmining
Rhodococcus sp.	-	-	-
Rhodococcus sp. AJ270	-	-	-

Oxidation Stability

Oxidation Stability	Organism
The enzyme is strongly inhibited by oxidizing agents like H2O2, ammonium persulfate and N-bromosuccinimide.	Rhodococcus sp.

Purification (Commentary)

Purification (Comment)	Organism
(NH4)2SO4 fractionation, ion-exchange/hydrophobic/gel-filtration chromatography, stabilization by n-butyric acid, yield of purified protein: 23.96%	Rhodococcus sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	no conversion of KCN	Rhodococcus sp.
829.5	-	purified protein	Rhodococcus sp.

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(1R,3R)-2,2-dibromo-3-phenylcyclopropanecarbonitrile + H2O	substrate conversion: 11.6%, enantiomeric excess: 83.8, 88.9 (methanol), 81.0 (n-hexane)	Rhodococcus sp.	(1R,3R)-2,2-dibromo-3-phenylcyclopropanecarbamide	-	r
(1R,3R)-2,2-dibromo-3-phenylcyclopropanecarbonitrile + H2O	substrate conversion: 11.6%, enantiomeric excess: 83.8, 88.9 (methanol), 81.0 (n-hexane)	Rhodococcus sp. AJ270	(1R,3R)-2,2-dibromo-3-phenylcyclopropanecarbamide	-	r
(1R,3R)-3-phenylcyclopropanecarbonitrile + H2O	substrate conversion: 49.1%, enantiomeric excess: 22.7, 31.1 (methanol), 21.6 (n-hexane)	Rhodococcus sp.	(1R,3R)-3-phenylcyclopropanecarbamide	-	r
(1R,3R)-3-phenylcyclopropanecarbonitrile + H2O	substrate conversion: 49.1%, enantiomeric excess: 22.7, 31.1 (methanol), 21.6 (n-hexane)	Rhodococcus sp. AJ270	(1R,3R)-3-phenylcyclopropanecarbamide	-	r
(1R,3S)-3-phenylcyclopropanecarbonitrile + H2O	substrate conversion: 25.8%, enantiomeric excess: 95.4, 95.4 (methanol), 95.5 (n-hexane)	Rhodococcus sp.	(1R,3S)-3-phenylcyclopropanecarbamide	-	r
(1R,3S)-3-phenylcyclopropanecarbonitrile + H2O	substrate conversion: 25.8%, enantiomeric excess: 95.4, 95.4 (methanol), 95.5 (n-hexane)	Rhodococcus sp. AJ270	(1R,3S)-3-phenylcyclopropanecarbamide	-	r
(1S,3S)-2,2-dimethyl-3-phenylcyclopropanecarbonitrile + H2O	substrate conversion: 40.3%, enantiomeric excess: 84.7	Rhodococcus sp.	(1S,3S)-2,2-dimethyl-3-phenylcyclopropanecarbamide	-	r
(1S,3S)-2,2-dimethyl-3-phenylcyclopropanecarbonitrile + H2O	substrate conversion: 7.9%, enantiomeric excess: 3.2, 5.9 (methanol), 0.7 (n-hexane)	Rhodococcus sp.	(1S,3S)-2,2-dimethyl-3-phenylcyclopropanecarbamide	-	r
(S)-3-benzoyloxypentanedinitrile + H2O	substrate conversion: 38.5%, enantiomeric excess: 68.2	Rhodococcus sp.	3-amino-1-(2-amino-2-oxoethyl)-3-oxopropyl benzoate	-	r
1-(4-bromo-phenyl)-aziridine-2-carbonitrile + H2O	-	Rhodococcus sp.	1-(4-bromophenyl)aziridine-2-carboxamide	-	r
1-(4-methoxy-phenyl)-aziridine-2-carbonitrile + H2O	-	Rhodococcus sp.	1-(4-methoxyphenyl)aziridine-2-carboxamide	-	r
2,3-dihydro-benzo(1,4)dioxine-2-carbonitrile + H2O	substrate conversion: 45.1%, enantiomeric excess: 0	Rhodococcus sp.	2,3-dihydro-1,4-benzodioxine-2-carboxamide	-	r
2-hydroxymethyl-3-phenyl-propionitrile + H2O	-	Rhodococcus sp.	2-benzyl-3-hydroxypropanamide	-	r
2-methoxymethyl-3-phenyl-propionitrile + H2O	-	Rhodococcus sp.	2-benzyl-3-methoxypropanamide	-	r
3-allyloxy-4-phenyl-butyronitrile + H2O	-	Rhodococcus sp.	4-phenyl-3-(prop-2-en-1-yloxy)butanamide	-	r
3-benzyloxy-3-vinyl-propionitrile + H2O	-	Rhodococcus sp.	3-(benzyloxy)pent-4-enamide	-	r
3-benzyloxy-pentanonitrile + H2O	-	Rhodococcus sp.	3-(benzyloxy)pentanamide	-	r
acrylonitrile + H2O	-	Rhodococcus sp.	acrylamide	-	r
acrylonitrile + H2O	-	Rhodococcus sp. AJ270	acrylamide	-	r
alpha-methylbenzyl cyanide + H2O	-	Rhodococcus sp.	2-phenylpropanamide	-	r
benzylcyanide + H2O	-	Rhodococcus sp.	2-phenylacetamide	-	r
n-butyronitrile + H2O	-	Rhodococcus sp.	n-butyramide	-	r
propionitrile + H2O	-	Rhodococcus sp.	propionamide	-	r
propionitrile + H2O	-	Rhodococcus sp. AJ270	propionamide	-	r

Subunits

Subunits	Comment	Organism
heterodimer	alphabeta, 1 * 22975 + 1 * 23493, MALDI-TOF MS	Rhodococcus sp.

Synonyms

Synonyms	Comment	Organism
NHase	-	Rhodococcus sp.
nitrile hydratase	-	Rhodococcus sp.

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	hydration of acrylonitrile	Rhodococcus sp.

pH Range

pH Minimum	pH Maximum	Comment	Organism
7.2	7.6	hydration of acrylonitrile at 25°C, rapid decrease of activity below pH 6.4	Rhodococcus sp.