Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.4.3.19 extracted from

  • Job, V.; Molla, G.; Pilone, M.S.; Pollegioni, L.
    Overexpression of a recombinant wild-type anf His-tagged Bacillus subtilis glycine oxidase in Escherichia coli (2002), Eur. J. Biochem., 269, 1456-1463.
    View publication on PubMed
Search for more literature for 1.4.3.19

Cloned(Commentary)

Cloned (Comment) Organism
production of a recombinant plasmid, pT7-GO by insertion of the DNA encoding for glycine oxidase into the multiple cloning site of the expression vector pT7.7. The pT7-glycine oxidase encodes a fully active fusion protein with six additional residues at the N-terminus of glycine oxidase. In BL21(DE3)pLysS Escherichia coli cells, and under optimal isopropyl thio-beta-D-galactoside induction conditions, soluble and active chimeric glycine oxidase is expressed up to 1.14 U per mg of cell. An N-terminally His-tagged glycine oxidase is also successfully expressed in Escherichia coli as a soluble protein and a fully active holoenzyme Bacillus subtilis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
46600
-
 4 * 46600, recombinant glycine oxidase, SDS-PAGE Bacillus subtilis
49400
-
 4 * 49400, recombinant His-tagged chimeric glycine oxidase Bacillus subtilis
166400
-
 recombinant His-tagged chimeric glycine oxidase, gel filtration Bacillus subtilis
187600
-
 recombinant glycine oxidase, gel filtration Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type enzyme and His-tagged chimeric glycine oxidase Bacillus subtilis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.568
-
 wild-type enzyme Bacillus subtilis
1.06
-
 His-tagged chimeric glycine oxidase Bacillus subtilis

Storage Stability

Storage Stability Organism
-80°C, pH 7.5, both glycine oxidase and His-tagged glycine oxidase are stable for months Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-2-aminobutyrate + H2O + O2 as active as sarcosine Bacillus subtilis 2-oxobutyrate + H2O2
-
 ?
D-Ala + H2O + O2 about 115% of the activity with sarcosine Bacillus subtilis pyruvate + NH3 + H2O2
-
 ?
D-Arg + H2O + O2 about 35% of the activity with sarcosine Bacillus subtilis ? + H2O2
-
 ?
D-His + H2O + O2 about 25% of the activity with sarcosine Bacillus subtilis ? + H2O2
-
 ?
D-Ile + H2O + O2 about 30% of the activity with sarcosine Bacillus subtilis 3-methyl-2-oxopentanoate + H2O2
-
 ?
D-Leu + H2O + O2 about 35% of the activity with sarcosine Bacillus subtilis 4-methyl-2-oxopentanoate + H2O2
-
 ?
D-pipecolate + H2O + O2 about 85% of the activity with sarcosine Bacillus subtilis ? + H2O2
-
 ?
D-Pro + H2O + O2 about 120% of the activity with sarcosine Bacillus subtilis ? + H2O2
-
 ?
D-Val + H2O + O2 about 35% of the activity with sarcosine Bacillus subtilis 3-methyl-2-oxobutanoate + H2O2
-
 ?
glycine + H2O + O2 as active as sarcosine Bacillus subtilis glyoxylate + NH3 + H2O2
-
 ?
additional information the enzyme is strictly stereospecific as it only catalyzes the oxidation of the D-isomer Bacillus subtilis ?
-
 ?
N-ethylglycine + H2O + O2 about 65% of the activity with sarcosine Bacillus subtilis glyoxylate + ethylamine + H2O2
-
 ?
N-methyl-D-Ala + H2O + O2 about 110% of the activity with sarcosine Bacillus subtilis pyruvate + methylamine + H2O2
-
 ?
sarcosine + H2O + O2
-
 Bacillus subtilis glyoxylate + methylamine + H2O2
-
 ?

Subunits

Subunits Comment Organism
tetramer 4 * 46600, recombinant glycine oxidase, SDS-PAGE Bacillus subtilis
tetramer 4 * 49400, recombinant His-tagged chimeric glycine oxidase Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
additional information
-
 increase in activity with no evidence for any plateau or decrease up to 60°C, reaction with sarcosine Bacillus subtilis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
15 60 increase in activity with no evidence for any plateau or decrease up to 60°C, reaction with sarcosine Bacillus subtilis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
35
-
 pH 7.0, 30 min, stable up to Bacillus subtilis
45
-
 pH 7.0, 30 min, about 30% loss of activity Bacillus subtilis
46
-
 Tm-value, after 30 min Bacillus subtilis
50
-
 pH 7.0, 30 min, about 75% loss of activity Bacillus subtilis
60
-
 pH 7.0, 30 min, complete loss of activity Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10
-
 reaction with sarcosine Bacillus subtilis

pH Range

pH Minimum pH Maximum Comment Organism
8 11 pH 8.0: about 55% of maximal activity, pH 11.0: about 55% of maximal activity, reaction with sarcosine Bacillus subtilis

pH Stability

pH Stability pH Stability Maximum Comment Organism
7 8.5 25°C, 300 min, maximal stability Bacillus subtilis

Cofactor

Cofactor Comment Organism Structure
FAD tight binding of coenzyme to the protein moiety, addition of exogenous FAD or FMN to the assay mixture does not increase enzyme activity Bacillus subtilis

pI Value

Organism Comment pI Value Maximum pI Value
Bacillus subtilis theoretical value, His-tagged chimeric enzyme
-
 6
Bacillus subtilis theoretical value, wild-type enzyme
-
 6