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Literature summary for 1.4.3.19 extracted from

  • Tatsumi, M.; Hoshino, W.; Kodama, Y.; Ueatrongchit, T.; Takahashi, K.; Yamaguchi, H.; Tagami, U.; Miyano, H.; Asano, Y.; Mizukoshi, T.
    Development of a rapid and simple glycine analysis method using a stable glycine oxidase mutant (2019), Anal. Biochem., 587, 113447 .
    View publication on PubMed
Search for more literature for 1.4.3.19

Application

Application Comment Organism
diagnostics a rapid and simple glycine analysis method is developed using a stable glycine oxidase mutant (T42 A/C245 S/L301V). Glycine analysis is important in research fields such as physiology and healthcare because the concentration of glycine in human plasma has been reported to change with various disorders Bacillus subtilis

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21 (DE3) Bacillus subtilis

Protein Variants

Protein Variants Comment Organism
L301V improvement in residual activity after incubation of 60 min at 60°C to 12.9% Bacillus subtilis
T42A improvement in residual activity after incubation of 60 min at 60°C to 18.7% Bacillus subtilis
T42A/C245S/L301V improvement in residual activity after incubation of 60 min at 60°C to 74.0%. The mutant enzyme retains most of its enzymatic activity during storage for over a year at 4°C Bacillus subtilis
T42S improvement in residual activity after incubation of 60 min at 60°C to 31.5% Bacillus subtilis
T42S/C245S/L301V improvement in residual activity after incubation of 60 min at 60°C to 55.7% Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.047
-
 glycine pH 8.5, 37°C, mutant enzyme T42A/C245S/L301V Bacillus subtilis
0.99
-
 glycine pH 8.5, 37°C, wild-type enzyme Bacillus subtilis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
glycine + H2O + O2 Bacillus subtilis
-
 glyoxylate + NH3 + H2O2
-
 ?
glycine + H2O + O2 Bacillus subtilis 168
-
 glyoxylate + NH3 + H2O2
-
 ?

Organism

Organism UniProt Comment Textmining
Bacillus subtilis O31616
-
-
Bacillus subtilis 168 O31616
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
glycine + H2O + O2
-
 Bacillus subtilis glyoxylate + NH3 + H2O2
-
 ?
glycine + H2O + O2
-
 Bacillus subtilis 168 glyoxylate + NH3 + H2O2
-
 ?

Synonyms

Synonyms Comment Organism
GlyOX
-
 Bacillus subtilis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
 60 min, residual activity of wild-type is almost absent, activity of C245S mutant is 4.4% Bacillus subtilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.3
-
 glycine pH 8.5, 37°C, mutant enzyme T42A/C245S/L301V Bacillus subtilis
1.3
-
 glycine pH 8.5, 37°C, wild-type enzyme Bacillus subtilis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.3
-
 glycine pH 8.5, 37°C, wild-type enzyme Bacillus subtilis
6.4
-
 glycine pH 8.5, 37°C, mutant enzyme T42A/C245S/L301V Bacillus subtilis