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Literature summary for 1.10.3.15 extracted from

  • Suzuki, H.; Furusho, Y.; Higashi, T.; Ohnishi, Y.; Horinouchi, S.
    A novel o-aminophenol oxidase responsible for formation of the phenoxazinone chromophore of grixazone (2006), J. Biol. Chem., 281, 824-833.
    View publication on PubMed
Search for more literature for 1.10.3.15

Activating Compound

Activating Compound Comment Organism Structure
GriE protein GriE activates the enzyme by transferring copper ions Streptomyces griseus

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3)/pLysS cells Streptomyces griseus

Inhibitors

Inhibitors Comment Organism Structure
2-Nitrophenol
-
 Streptomyces griseus
3-amino-4-hydroxybenzensulfonic acid
-
 Streptomyces griseus
4-hydroxy-3-nitrobenzaldehyde
-
 Streptomyces griseus
4-hydroxybenzaldehyde
-
 Streptomyces griseus
aniline
-
 Streptomyces griseus
L-tyrosine
-
 Streptomyces griseus
additional information 10 mM EDTA and o-phenanthroline have negligible effects on the purified enzyme Streptomyces griseus
phenol
-
 Streptomyces griseus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.41
-
 3,4-Dihydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
0.58
-
 3-amino-4-hydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
0.75
-
 2-amino-4-methylphenol at pH 7.0 and 55°C Streptomyces griseus
3.5
-
 o-aminophenol at pH 7.0 and 55°C Streptomyces griseus
5.5
-
 L-Dopa at pH 7.0 and 55°C Streptomyces griseus
19
-
 catechol at pH 7.0 and 55°C Streptomyces griseus

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ the enzyme contains two copper ions in the active site. 10 mM Cu2+ specifically increases the enzyme activity by about 20fold Streptomyces griseus
additional information Zn2+, Ni2+, Co2+, Mn2+, Fe2+, or Mg2+ exert almost no effects Streptomyces griseus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36000
-
 x * 36000, SDS-PAGE Streptomyces griseus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3-amino-4-hydroxybenzoate + N-acetyl-L-cysteine + O2 Streptomyces griseus
-
 grixazone B + H2O + CO2
-
 ?

Organism

Organism UniProt Comment Textmining
Streptomyces griseus
-
-
-
Streptomyces griseus IFO 13350
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Cu2+-charged HiTrap chelating column chromatography, HiTrap Q column chromatography, Resource PHE column chromatography, and Superdex 200 gel filtration Streptomyces griseus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0000035
-
 crude extract, at pH 7.0 and 30°C Streptomyces griseus
0.0658
-
 after 19000fold purification, at pH 7.0 and 30°C Streptomyces griseus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-amino-4-methylphenol + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
3,4-dihydroxybenzaldehyde + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
3-amino-4-hydroxybenzaldehyde + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus grixazone A + H2O + CO2
-
 ?
3-amino-4-hydroxybenzoate + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus grixazone B + H2O + CO2
-
 ?
4-amino-3-hydroxybenzoate + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
catechol + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
L-Dopa + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
additional information the enzyme shows no monophenolase activity, although it oxidizes various o-aminophenols as preferable substrates rather than catechol-type substrates. The enzyme does not use tyrosine as a substrate Streptomyces griseus ?
-
 ?
o-aminophenol + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus 2-aminophenoxazin-3-one + H2O + CO2
-
 ?

Subunits

Subunits Comment Organism
? x * 36000, SDS-PAGE Streptomyces griseus

Synonyms

Synonyms Comment Organism
GriF
-
 Streptomyces griseus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
 at pH 7.0 Streptomyces griseus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 70 more than 50% activity between 30 and 70°C Streptomyces griseus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
10 55 when the enzyme sample is kept at pH 7.0 for 20 min, it remains active below 55°C, and activity drops sharply above 55°C Streptomyces griseus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.066
-
 L-Dopa at pH 7.0 and 55°C Streptomyces griseus
0.8
-
 3,4-Dihydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
12
-
 catechol at pH 7.0 and 55°C Streptomyces griseus
14
-
 3-amino-4-hydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
18
-
 2-amino-4-methylphenol at pH 7.0 and 55°C Streptomyces griseus
20
-
 o-aminophenol at pH 7.0 and 55°C Streptomyces griseus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5 10.5
-
 Streptomyces griseus

pH Range

pH Minimum pH Maximum Comment Organism
6 11 more than 50% activity between pH 6.0 and 11.0 Streptomyces griseus

pH Stability

pH Stability pH Stability Maximum Comment Organism
6.5 8.5 the enzyme is most stable at pH 6.5-8.5 Streptomyces griseus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.9
-
 4-hydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
3.2
-
 L-tyrosine at pH 7.0 and 55°C Streptomyces griseus
3.9
-
 4-hydroxy-3-nitrobenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
6.1
-
 2-Nitrophenol at pH 7.0 and 55°C Streptomyces griseus
14
-
 aniline at pH 7.0 and 55°C Streptomyces griseus
21
-
 phenol at pH 7.0 and 55°C Streptomyces griseus
31
-
 3-amino-4-hydroxybenzensulfonic acid at pH 7.0 and 55°C Streptomyces griseus

General Information

General Information Comment Organism
metabolism GriF is an o-aminophenol oxidase responsible for the in vivo formation of the phenoxazinone chromophore in the grixazone biosynthetic pathway Streptomyces griseus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0073
-
 4-Amino-3-hydroxybenzoate at pH 7.0 and 55°C Streptomyces griseus
0.012
-
 L-Dopa at pH 7.0 and 55°C Streptomyces griseus
0.19
-
 3-amino-4-hydroxybenzoate at pH 7.0 and 55°C Streptomyces griseus
0.32
-
 catechol at pH 7.0 and 55°C Streptomyces griseus
0.97
-
 3,4-Dihydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
5.8
-
 o-aminophenol at pH 7.0 and 55°C Streptomyces griseus
23
-
 2-amino-4-methylphenol at pH 7.0 and 55°C Streptomyces griseus
24
-
 3-amino-4-hydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus