Literature summary for 4.2.1.84 extracted from

Cheng, Z.; Peplowski, L.; Cui, W.; Xia, Y.; Liu, Z.; Zhang, J.; Kobayashi, M.; Zhou, Z.
Identification of key residues modulating the stereoselectivity of nitrile hydratase toward rac-mandelonitrile by semi-rational engineering (2018), Biotechnol. Bioeng., 115, 524-535 .

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Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Rhodococcus rhodochrous

Protein Variants

Protein Variants	Comment	Organism
A51L	wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 74.3%	Rhodococcus rhodochrous
F37H	wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 96.8%	Rhodococcus rhodochrous
F37H/F51L	wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 94.6%	Rhodococcus rhodochrous
F37H/L48A	wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 95.1%	Rhodococcus rhodochrous
F37Y	wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 88.7%	Rhodococcus rhodochrous
F51Q	wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 60.2%	Rhodococcus rhodochrous
L48Q	wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 59.7%	Rhodococcus rhodochrous

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
38.2	-	(S)-mandelonitrile	pH 7.4, 10°C, wild-type enzyme	Rhodococcus rhodochrous
41.9	-	(S)-mandelonitrile	pH 7.4, 10°C, mutant enzyme F37H	Rhodococcus rhodochrous
43.8	-	(R)-mandelonitrile	pH 7.4, 10°C, wild-type enzyme	Rhodococcus rhodochrous

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
(R)-mandelonitrile + H2O	Rhodococcus rhodochrous	wild-type enzyme catalyzes the conversion of rac-mandelonitrile to (S)-mandelamide with an enantiomeric excess of 52.6%	(R)-mandelamide	-	?
(R)-mandelonitrile + H2O	Rhodococcus rhodochrous J1	wild-type enzyme catalyzes the conversion of rac-mandelonitrile to (S)-mandelamide with an enantiomeric excess of 52.6%	(R)-mandelamide	-	?
(S)-mandelonitrile + H2O	Rhodococcus rhodochrous	wild-type enzyme catalyzes the conversion of rac-mandelonitrile to (S)-mandelamide with an enantiomeric excess of 52.6%	(S)-mandelamide	-	?
(S)-mandelonitrile + H2O	Rhodococcus rhodochrous J1	wild-type enzyme catalyzes the conversion of rac-mandelonitrile to (S)-mandelamide with an enantiomeric excess of 52.6%	(S)-mandelamide	-	?

Organism

Organism	UniProt	Comment	Textmining
Rhodococcus rhodochrous	P29378 AND P29379	P29378: alpha-subunit, P29379: beta-subunit	-
Rhodococcus rhodochrous J1	P29378 AND P29379	P29378: alpha-subunit, P29379: beta-subunit	-

Purification (Commentary)

Purification (Comment)	Organism
-	Rhodococcus rhodochrous

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(R)-mandelonitrile + H2O	wild-type enzyme catalyzes the conversion of rac-mandelonitrile to (S)-mandelamide with an enantiomeric excess of 52.6%	Rhodococcus rhodochrous	(R)-mandelamide	-	?
(R)-mandelonitrile + H2O	wild-type enzyme catalyzes the conversion of rac-mandelonitrile to (S)-mandelamide with an enantiomeric excess of 52.6%	Rhodococcus rhodochrous J1	(R)-mandelamide	-	?
(S)-mandelonitrile + H2O	wild-type enzyme catalyzes the conversion of rac-mandelonitrile to (S)-mandelamide with an enantiomeric excess of 52.6%	Rhodococcus rhodochrous	(S)-mandelamide	-	?
(S)-mandelonitrile + H2O	wild-type enzyme catalyzes the conversion of rac-mandelonitrile to (S)-mandelamide with an enantiomeric excess of 52.6%	Rhodococcus rhodochrous J1	(S)-mandelamide	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.74	-	(R)-mandelonitrile	pH 7.4, 10°C, wild-type enzyme	Rhodococcus rhodochrous
1.2	-	(S)-mandelonitrile	pH 7.4, 10°C, wild-type enzyme	Rhodococcus rhodochrous
1.4	-	(S)-mandelonitrile	pH 7.4, 10°C, mutant enzyme F37H	Rhodococcus rhodochrous

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.0168	-	(R)-mandelonitrile	pH 7.4, 10°C, wild-type enzyme	Rhodococcus rhodochrous
0.032	-	(S)-mandelonitrile	pH 7.4, 10°C, wild-type enzyme	Rhodococcus rhodochrous
0.034	-	(S)-mandelonitrile	pH 7.4, 10°C, mutant enzyme F37H	Rhodococcus rhodochrous

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Release 2023.1 (January 2023)