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Literature summary for 1.7.3.3 extracted from

  • Yainoy, S.; Phuadraksa, T.; Wichit, S.; Sompoppokakul, M.; Songtawee, N.; Prachayasittikul, V.; Isarankura-Na-Ayudhya, C.
    Production and characterization of recombinant wild type uricase from indonesian coelacanth (L. menadoensis) and improvement of its thermostability by in silico rational design and disulphide bridges engineering (2019), Int. J. Mol. Sci., 20, 1269 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.7.3.3

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Latimeria menadoensis

Protein Variants

Protein Variants Comment Organism
A132C/A225C the mutant shows reduced catalytic efficiency compared to the wild type enzyme Latimeria menadoensis
D288C the mutant shows 0.8fold reduced catalytic efficiency compared to the wild type enzyme Latimeria menadoensis
I27C/N289C the mutant with 0.45fold reduced catalytic efficiency has similar specific activity and production yield to that of wild type but its thermostability is dramatically improved (120fold increase in half-life at 37°C) Latimeria menadoensis
N289C the mutant 0.3fold reduced catalytic efficiency compared to the wild type enzyme Latimeria menadoensis
S136C the mutant shows reduced catalytic efficiency compared to the wild type enzyme Latimeria menadoensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.01528
-
 Urate mutant enzyme S136C, at pH 8.0 and 30°C Latimeria menadoensis
0.01843
-
 Urate wild type enzyme, at pH 8.0 and 30°C Latimeria menadoensis
0.03046
-
 Urate mutant enzyme N289C, at pH 8.0 and 30°C Latimeria menadoensis
0.03539
-
 Urate mutant enzyme I27C/N289C, at pH 8.0 and 30°C Latimeria menadoensis
0.03919
-
 Urate mutant enzyme A132C/A225C, at pH 8.0 and 30°C Latimeria menadoensis
0.18965
-
 Urate mutant enzyme D288C, at pH 8.0 and 30°C Latimeria menadoensis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
150000
-
 gel filtration Latimeria menadoensis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
urate + O2 + H2O Latimeria menadoensis
-
 allantoin + H2O2
-
 ?

Organism

Organism UniProt Comment Textmining
Latimeria menadoensis U4KJC3
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography and Sephacryl S-300 gel filtration Latimeria menadoensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
7.88
-
 mutant enzyme S136C, at pH 8.0 and 30°C Latimeria menadoensis
9.38
-
 mutant enzyme D288C, at pH 8.0 and 30°C Latimeria menadoensis
9.75
-
 mutant enzyme I27C/N289C, at pH 8.0 and 30°C Latimeria menadoensis
10.38
-
 mutant enzyme N289C, at pH 8.0 and 30°C Latimeria menadoensis
10.45
-
 wild type enzyme, at pH 8.0 and 30°C Latimeria menadoensis
15.38
-
 mutant enzyme A132C/A225C, at pH 8.0 and 30°C Latimeria menadoensis

Storage Stability

Storage Stability Organism
-20°C and 4°C, mutant enzyme I27C/N289C, 60 days, almost no loss of activity Latimeria menadoensis
37°C, mutant enzyme I27C/N289C, 15 days, almost no loss of activity Latimeria menadoensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
urate + O2 + H2O
-
 Latimeria menadoensis allantoin + H2O2
-
 ?

Subunits

Subunits Comment Organism
homotetramer 4 * 36400, calculated from amino acid sequence Latimeria menadoensis

Synonyms

Synonyms Comment Organism
Uox
-
 Latimeria menadoensis
uricase
-
 Latimeria menadoensis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
-
 Latimeria menadoensis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
25 50 more than 50% activity between 25 and 50°C Latimeria menadoensis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 the enzyme retains almost 100% activity after 2 min at 45°C, however, at higher temperatures the activity is rapidly lost Latimeria menadoensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
22.49
-
 Urate mutant enzyme S136C, at pH 8.0 and 30°C Latimeria menadoensis
29.89
-
 Urate wild type enzyme, at pH 8.0 and 30°C Latimeria menadoensis
32.58
-
 Urate mutant enzyme I27C/N289C, at pH 8.0 and 30°C Latimeria menadoensis
34.06
-
 Urate mutant enzyme N289C, at pH 8.0 and 30°C Latimeria menadoensis
58.19
-
 Urate mutant enzyme A132C/A225C, at pH 8.0 and 30°C Latimeria menadoensis
65.07
-
 Urate mutant enzyme D288C, at pH 8.0 and 30°C Latimeria menadoensis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Latimeria menadoensis

pH Range

pH Minimum pH Maximum Comment Organism
7 11 more than 50% activity between pH 7.0 and 11.0 Latimeria menadoensis

Cofactor

Cofactor Comment Organism Structure
additional information the enzyme is cofactor-free Latimeria menadoensis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
340
-
 Urate mutant enzyme D288C, at pH 8.0 and 30°C Latimeria menadoensis
920
-
 Urate mutant enzyme I27C/N289C, at pH 8.0 and 30°C Latimeria menadoensis
1120
-
 Urate mutant enzyme N289C, at pH 8.0 and 30°C Latimeria menadoensis
1470
-
 Urate mutant enzyme S136C, at pH 8.0 and 30°C Latimeria menadoensis
1490
-
 Urate mutant enzyme A132C/A225C, at pH 8.0 and 30°C Latimeria menadoensis
1620
-
 Urate wild type enzyme, at pH 8.0 and 30°C Latimeria menadoensis