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Literature summary for 3.5.3.6 extracted from

  • Cai, X.; Jiang, H.; Zhang, T.; Jiang, B.; Mu, W.; Miao, M.
    Thermostability and specific-activity enhancement of an arginine deiminase from Enterococcus faecalis SK23.001 via semirational design for L-citrulline production (2018), J. Agric. Food Chem., 66, 8841-8850 .
    View publication on PubMed
Search for more literature for 3.5.3.6

Application

Application Comment Organism
synthesis L-citrulline can be enzymatically produced by arginine deiminase Enterococcus faecalis

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21(DE3) Enterococcus faecalis

Protein Variants

Protein Variants Comment Organism
F269P the conversion rate of L-arginine to L-citrulline at 45°C is 93.2% compared to wild-type enzyme Enterococcus faecalis
G292C the conversion rate of L-arginine to L-citrulline at 45°C is 96.0% compared to wild-type enzyme Enterococcus faecalis
G292D the conversion rate of L-arginine to L-citrulline at 45°C is 97.5% compared to wild-type enzyme Enterococcus faecalis
G292P the conversion rate of L-arginine to L-citrulline at 45°C is 96.5% compared to wild-type enzyme Enterococcus faecalis
R15K the conversion rate of L-arginine to L-citrulline at 45°C is 95.8% compared to wild-type enzyme Enterococcus faecalis
R15K/F269Y/G292P the triple-site mutant enzyme displays a 2.5fold higher specific enzyme activity (333 U/mg), a lower Km value of 6.4 mM, and a 6.1fold longer half-life (t1/2(45°C): 86.7 min) than wild-type enzyme. The conversion rate of L-arginine to L-citrulline at 45°C is 97.4% compared to wild-type enzyme Enterococcus faecalis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.4
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/F269Y/G292P Enterococcus faecalis
6.7
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/G292P Enterococcus faecalis
9.3
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/F269Y Enterococcus faecalis
10.1
-
 L-arginine pH 6.0, 45°C, wild-type enzyme Enterococcus faecalis
11.6
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K Enterococcus faecalis
12.5
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/G264P Enterococcus faecalis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-arginine + H2O Enterococcus faecalis
-
 L-citrulline + NH3
-
 ?
L-arginine + H2O Enterococcus faecalis SK23.001
-
 L-citrulline + NH3
-
 ?

Organism

Organism UniProt Comment Textmining
Enterococcus faecalis A0A3G2YMH7
-
-
Enterococcus faecalis SK23.001 A0A3G2YMH7
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
126.5
-
 substrate: L-arginine, pH 6.0, 45°C, mutant enzyme R15K/F269Y Enterococcus faecalis
131.2
-
 substrate: L-arginine, pH 6.0, 45°C, wild-type enzyme Enterococcus faecalis
134.7
-
 substrate: L-arginine, pH 6.0, 45°C, mutant enzyme R15K/G264P Enterococcus faecalis
143.2
-
 substrate: L-arginine, pH 6.0, 45°C, mutant enzyme R15K Enterococcus faecalis
296.8
-
 substrate: L-arginine, pH 6.0, 45°C, mutant enzyme R15K/G292P Enterococcus faecalis
332.8
-
 substrate: L-arginine, pH 6.0, 45°C, mutant enzyme R15K/F269Y/G292P Enterococcus faecalis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-arginine + H2O
-
 Enterococcus faecalis L-citrulline + NH3
-
 ?
L-arginine + H2O
-
 Enterococcus faecalis SK23.001 L-citrulline + NH3
-
 ?

Synonyms

Synonyms Comment Organism
ADI
-
 Enterococcus faecalis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
 Enterococcus faecalis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 t1/2: 14.3 (wild-type enzyme), t1/2: 14.5 (mutant enzyme R15K), t1/2: 14.9 (mutant enzyme R15K/G264P), t1/2: 48.0 (mutant enzyme R15K/F269Y), t1/2: 82.5 (mutant enzyme R15K/G292P), t1/2: 86.7 min (mutant enzyme R15K/F269Y/G292P) Enterococcus faecalis
60
-
 t1/2: 12 (wild-type enzyme) Enterococcus faecalis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
268
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K Enterococcus faecalis
291
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/G264P Enterococcus faecalis
309
-
 L-arginine pH 6.0, 45°C, wild-type enzyme Enterococcus faecalis
342
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/F269Y Enterococcus faecalis
365
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/F269Y/G292P Enterococcus faecalis
371
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/G292P Enterococcus faecalis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Enterococcus faecalis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
23.1
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K Enterococcus faecalis
23.3
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/G264P Enterococcus faecalis
30.6
-
 L-arginine pH 6.0, 45°C, wild-type enzyme Enterococcus faecalis
36.8
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/F269Y Enterococcus faecalis
55.4
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/G292P Enterococcus faecalis
57
-
 L-arginine pH 6.0, 45°C, mutant enzyme R15K/F269Y/G292P Enterococcus faecalis