Literature summary for 3.5.3.6 extracted from

El-Sayed, A.; Hassan, M.; Nada, H.
Purification, immobilization, and biochemical characterization of L-arginine deiminase from thermophilic Aspergillus fumigatus KJ434941 Anticancer activity in vitro (2015), Biotechnol. Prog., 31, 396-405 .

Search for more literature for 3.5.3.6

Application

Application	Comment	Organism
drug development	L-arginine deiminase has a powerful anticancer activity against various tumors, via arginine depletion, arresting the cell cycle at G1 phase. The free and PEGylated enzyme exhibits a similar cytotoxic efficacy against HCT, HEP-G2, and MCF7 cells, lower than the cytotoxic efficacy of to enzyme covalently immobilized on dextran. The in vitro anticancer activity of the enzyme against HCT, MCF7, and HEPG-2 cells is increased by five-, three-, and threefold upon covalent modification by dextran	Aspergillus fumigatus

General Stability

General Stability	Organism
upon proteolysis for 30 min, the residual activity of the soluble enzyme, the PEGylated enzyme, and the enzyme covalently immobilized on dextran is 8.0%, 32.0%, and 20.0% for proteinase K and 10.0%, 52.0%, and 90.0% for acid protease, respectively	Aspergillus fumigatus

Inhibitors

Inhibitors	Comment	Organism	Structure
6-diazo-5-oxo-norleucine	-	Aspergillus fumigatus
hydroxylamine	-	Aspergillus fumigatus
iodoacetate	-	Aspergillus fumigatus
propargylglycine	-	Aspergillus fumigatus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
8.66	-	L-alanine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
8.76	-	L-arginine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
12.34	-	L-lysine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
15.48	-	L-lysine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
15.48	-	L-alanine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
15.57	-	L-arginine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
18.42	-	L-lysine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
22.52	-	L-arginine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
23.21	-	L-alanine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
100000	110000	gel filtration	Aspergillus fumigatus

Organism

Organism	UniProt	Comment	Textmining
Aspergillus fumigatus	-	-	-
Aspergillus fumigatus KJ434941	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Aspergillus fumigatus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
18	-	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
21.5	-	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
26.7	-	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-alanine + H2O	-	Aspergillus fumigatus	? + NH3	-	?
L-alanine + H2O	-	Aspergillus fumigatus KJ434941	? + NH3	-	?
L-arginine + H2O	-	Aspergillus fumigatus	L-citrulline + NH3	-	?
L-arginine + H2O	-	Aspergillus fumigatus KJ434941	L-citrulline + NH3	-	?
L-lysine + H2O	-	Aspergillus fumigatus	? + NH3	-	?
L-lysine + H2O	-	Aspergillus fumigatus KJ434941	? + NH3	-	?

Subunits

Subunits	Comment	Organism
homodimer	2 * 50000, SDS-PAGE	Aspergillus fumigatus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
40	-	soluble enzyme (t1/2: 12.1 h), EGylated enzyme (t1/2: 14.6 h), enzyme covalently immobilized on dextran (13.9 h)	Aspergillus fumigatus
50	-	PEGylated enzyme displays twofold resistance to thermal denaturation (t1/2: 13.9 h) compared to soluble enzyme (t1/2: 8.3 h)	Aspergillus fumigatus
60	-	soluble enzyme (t1/2: 5.7 h), EGylated enzyme (t1/2: 12.1 h), enzyme covalently immobilized on dextran (12.1 h)	Aspergillus fumigatus
70	-	thermal stability of PEGylated enzyme (9.1 h) is increased by 1.7fold. Similar stability to enzyme covalently immobilized on dextran (5.7 h) with the soluble enzyme (5.6 h)	Aspergillus fumigatus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.0024	-	L-alanine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
0.0033	-	L-lysine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
0.0034	-	L-alanine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
0.004	-	L-lysine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
0.0041	-	L-arginine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
0.0043	-	L-lysine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
0.00475	-	L-arginine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
0.00478	-	L-arginine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
0.006	-	L-alanine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.343	-	hydroxylamine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
0.387	-	iodoacetate	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
0.4511	-	iodoacetate	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
0.481	-	propargylglycine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
0.615	-	hydroxylamine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
0.783	-	propargylglycine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
1.164	-	hydroxylamine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
1.215	-	6-diazo-5-oxo-norleucine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
2.081	-	6-diazo-5-oxo-norleucine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
2.232	-	6-diazo-5-oxo-norleucine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
2.291	-	iodoacetate	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
2.739	-	propargylglycine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
1.9	-	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus	iodoacetate
1.98	-	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus	hydroxylamine
2.3	-	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus	hydroxylamine
2.52	-	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus	propargylglycine
2.6	-	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus	iodoacetate
3.22	-	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus	propargylglycine
4.9	-	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus	hydroxylamine
6.7	-	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus	6-diazo-5-oxo-norleucine
7.39	-	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus	iodoacetate
8.15	-	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus	6-diazo-5-oxo-norleucine
9.4	-	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus	6-diazo-5-oxo-norleucine
11.54	-	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus	propargylglycine

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.00018	-	L-arginine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
0.000215	-	L-lysine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
0.000215	-	L-alanine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
0.000235	-	L-lysine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
0.000258	-	L-alanine	pH 7.0, 45°C, enzyme covalently immobilized on dextran	Aspergillus fumigatus
0.000277	-	L-alanine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
0.0003	-	L-arginine	pH 7.0, 45°C, PEGylated enzyme	Aspergillus fumigatus
0.00032	-	L-lysine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus
0.00055	-	L-arginine	pH 7.0, 45°C, soluble enzyme	Aspergillus fumigatus

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Release 2023.1 (January 2023)