Literature summary for 4.1.2.47 extracted from

Asano, Y.; Dadashipour, M.; Yamazaki, M.; Doi, N.; Komeda, H.
Functional expression of a plant hydroxynitrile lyase in Escherichia coli by directed evolution: creation and characterization of highly in vivo soluble mutants (2011), Protein Eng. Des. Sel., 24, 607-616.

Search for more literature for 4.1.2.47

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli JM109 and BL21 (DE3) cells	Manihot esculenta

Protein Variants

Protein Variants	Comment	Organism
H103C	the mutant displays 9.3fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
H103I	the mutant displays 8.1fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
H103K	inactive	Manihot esculenta
H103L	the mutant displays 11.1fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
H103M	the mutant displays 9.07fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
H103P	inactive	Manihot esculenta
H103Q	the mutant displays 4.06fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
H103R	inactive	Manihot esculenta
H103S	the mutant displays 2.9fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
H103T	the mutant displays 3.4fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
H103W	inactive	Manihot esculenta
H103Y	inactive with (S)-mandelonitrile as substrate	Manihot esculenta
K176P	the mutant displays 2.02fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
K176P/K199P/K224P	the mutant displays 6.97fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
K176P/K224P	the mutant displays 5.05fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
K199P	the mutant displays 1.38fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
K199P/K224P	the mutant displays 4.25fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta
K224P	the mutant displays 2.53fold increase in total specific activity in the cell-free extract compared with the wild type	Manihot esculenta

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
1.86	-	(S)-mandelonitrile	mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
2	3	benzaldehyde	wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
2.5	5	(S)-mandelonitrile	mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
5.17	-	(S)-mandelonitrile	wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
5.54	-	(S)-mandelonitrile	mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
6.98	-	(S)-mandelonitrile	mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
12.3	-	benzaldehyde	mutant enzyme H103Y, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
13.4	-	benzaldehyde	mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
21.6	-	benzaldehyde	mutant enzyme K176P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
27	-	benzaldehyde	mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
27.9	-	benzaldehyde	mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta

Organism

Organism	UniProt	Comment	Textmining
Manihot esculenta	P52705	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(S)-mandelonitrile	-	Manihot esculenta	cyanide + benzaldehyde	-	r
cyanide + benzaldehyde	-	Manihot esculenta	(S)-mandelonitrile	-	r

Synonyms

Synonyms	Comment	Organism
HNL	-	Manihot esculenta
S-hydroxynitrile lyase	-	Manihot esculenta

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
21.5	-	(S)-mandelonitrile	mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
23.9	-	(S)-mandelonitrile	mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
34.5	-	(S)-mandelonitrile	wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
39.7	-	(S)-mandelonitrile	mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
41.8	-	(S)-mandelonitrile	mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
47.4	-	benzaldehyde	mutant enzyme H103Y, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
83.2	-	benzaldehyde	mutant enzyme K176P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
90.4	-	benzaldehyde	mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
93.5	-	benzaldehyde	mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
96.1	-	benzaldehyde	mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
96.6	-	benzaldehyde	wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta

Cofactor

Cofactor	Comment	Organism	Structure
FAD	-	Manihot esculenta

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
3.44	-	benzaldehyde	mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
3.46	-	benzaldehyde	mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
3.85	-	benzaldehyde	mutant enzyme H103Y, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
3.85	-	benzaldehyde	mutant enzyme K176P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
4.2	-	benzaldehyde	wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
6	-	(S)-mandelonitrile	mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
6.67	-	(S)-mandelonitrile	wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
6.75	-	benzaldehyde	mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication	Manihot esculenta
7.2	-	(S)-mandelonitrile	mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
8.43	-	(S)-mandelonitrile	mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta
12.8	-	(S)-mandelonitrile	mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication	Manihot esculenta

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Release 2023.1 (January 2023)