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Literature summary for 3.5.3.1 extracted from

  • Huang, K.; Mu, W.; Zhang, T.; Jiang, B.; Miao, M.
    Cloning, expression, and characterization of a thermostable L-arginase from Geobacillus thermodenitrificans NG80-2 for L-ornithine production (2016), Biotechnol. Appl. Biochem., 63, 391-397 .
    View publication on PubMed
Search for more literature for 3.5.3.1

Application

Application Comment Organism
synthesis the enzyme can be used to produce L-ornithine from L-arginine Geobacillus thermodenitrificans

Cloned(Commentary)

Cloned (Comment) Organism
overexpressed in Escherichia coli Geobacillus thermodenitrificans

Inhibitors

Inhibitors Comment Organism Structure
2-mercaptoethanol 1 mM, 12% loss of activity Geobacillus thermodenitrificans
5,5'-dithiobis (2-nitrobenzoic acid) 1 mM, 23% loss of activity Geobacillus thermodenitrificans
Ba2+ 2 mM, slight inhibition Geobacillus thermodenitrificans
Ca2+ 2 mM, slight inhibition Geobacillus thermodenitrificans
Cu2+ 2 mM, slight inhibition Geobacillus thermodenitrificans
diethylpyrocarbonate 1 mM, 83% loss of activity Geobacillus thermodenitrificans
Mg2+ 2 mM, slight inhibition Geobacillus thermodenitrificans
N-bromosuccinimide 1 mM, 93% loss of activity Geobacillus thermodenitrificans
Zn2+ 2 mM, slight inhibition Geobacillus thermodenitrificans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
171.9
-
 L-arginine pH 9.0, 80°C Geobacillus thermodenitrificans

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ enhances the activity Geobacillus thermodenitrificans
Mn2+ optimum metal cofactor. The optimal concentration of Mn2+ is approximately 2 mM. When the concentration is higher than 2 mM, there is a slight drop in relative activity. The active site of L-arginase contains two Mn2+ ions (Mn2+A and Mn2+B), which are coordinated by Asp and His residues. When Mn2+ concentration increases from 0 to 10 mM, the ellipticity of L-arginase at 208 nm and 222 nm increases accordingly. The percentage of helix increases to 66% in the presence of 10 mM Mn2+. This suggests that the addition of Mn2+ can affect the structure of L-arginase by increasing the helix content of the enzyme Geobacillus thermodenitrificans
Ni2+ enhances the activity Geobacillus thermodenitrificans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
192000
-
 gel filtration Geobacillus thermodenitrificans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-arginine + H2O Geobacillus thermodenitrificans
-
 L-ornithine + urea
-
 ?
L-arginine + H2O Geobacillus thermodenitrificans NG80-2
-
 L-ornithine + urea
-
 ?

Organism

Organism UniProt Comment Textmining
Geobacillus thermodenitrificans A4IJM8
-
-
Geobacillus thermodenitrificans NG80-2 A4IJM8
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Geobacillus thermodenitrificans

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
138.3
-
 pH 9.0, 80°C Geobacillus thermodenitrificans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-arginine + H2O
-
 Geobacillus thermodenitrificans L-ornithine + urea
-
 ?
L-arginine + H2O the biosynthesis yield of L-ornithine by the purified enzyme is 36.9 g/l, and the molar yield is 97.2% Geobacillus thermodenitrificans L-ornithine + urea
-
 ?
L-arginine + H2O
-
 Geobacillus thermodenitrificans NG80-2 L-ornithine + urea
-
 ?
L-arginine + H2O the biosynthesis yield of L-ornithine by the purified enzyme is 36.9 g/l, and the molar yield is 97.2% Geobacillus thermodenitrificans NG80-2 L-ornithine + urea
-
 ?

Subunits

Subunits Comment Organism
hexamer 6 * 33000, SDS-PAGE Geobacillus thermodenitrificans
hexamer 6 * 32300, calculated from sequence Geobacillus thermodenitrificans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
-
 Geobacillus thermodenitrificans

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 90 40°C: about 50% of maximal activity, 90°C: about 30% of maximal activity Geobacillus thermodenitrificans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
 20 min, enzyme activity retains 89.7% of its initial activity Geobacillus thermodenitrificans
50
-
 20 min, enzyme activity retains 87.1% of its initial activity Geobacillus thermodenitrificans
60
-
 20 min, enzyme activity retains 53.8% of its initial activity Geobacillus thermodenitrificans
70
-
 20 min, enzyme activity retains 52.1% of its initial activity Geobacillus thermodenitrificans
80
-
 20 min, enzyme activity retains 51.6% of its initial activity Geobacillus thermodenitrificans
90
-
 20 min, enzyme activity retains 30% of its initial activity Geobacillus thermodenitrificans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
657
-
 L-arginine pH 9.0, 80°C Geobacillus thermodenitrificans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Geobacillus thermodenitrificans

pH Range

pH Minimum pH Maximum Comment Organism
7 10 pH 7.0: about 40% of maximal activity, pH 10.0: about 75% of maximal activity Geobacillus thermodenitrificans

pI Value

Organism Comment pI Value Maximum pI Value
Geobacillus thermodenitrificans calculated
-
 5.49

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.8
-
 L-arginine pH 9.0, 80°C Geobacillus thermodenitrificans