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Literature summary 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

Activating Compound

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

Cloned(Commentary)

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.10.3.4 3-amino-4-hydroxybenzensulfonic acid
-
 Streptomyces griseus
1.10.3.4 4-hydroxy-3-nitrobenzaldehyde
-
 Streptomyces griseus
1.10.3.4 aniline
-
 Streptomyces griseus
1.10.3.4 L-tyrosine
-
 Streptomyces griseus
1.10.3.4 o-Nitrophenol
-
 Streptomyces griseus
1.10.3.4 p-Hydroxybenzaldehyde
-
 Streptomyces griseus
1.10.3.4 phenol
-
 Streptomyces griseus
1.10.3.15 2-Nitrophenol
-
 Streptomyces griseus
1.10.3.15 3-amino-4-hydroxybenzensulfonic acid
-
 Streptomyces griseus
1.10.3.15 4-hydroxy-3-nitrobenzaldehyde
-
 Streptomyces griseus
1.10.3.15 4-hydroxybenzaldehyde
-
 Streptomyces griseus
1.10.3.15 aniline
-
 Streptomyces griseus
1.10.3.15 L-tyrosine
-
 Streptomyces griseus
1.10.3.15 additional information 10 mM EDTA and o-phenanthroline have negligible effects on the purified enzyme Streptomyces griseus
1.10.3.15 phenol
-
 Streptomyces griseus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.10.3.4 0.41
-
 3,4-Dihydroxybenzaldehyde in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 0.58
-
 3-amino-4-hydroxybenzaldehyde in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 0.75
-
 2-amino-4-methylphenol in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 3.5
-
 o-aminophenol in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 5.5
-
 L-Dopa in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 19
-
 catechol in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.15 0.41
-
 3,4-Dihydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 0.58
-
 3-amino-4-hydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 0.75
-
 2-amino-4-methylphenol at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 3.5
-
 o-aminophenol at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 5.5
-
 L-Dopa at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 19
-
 catechol at pH 7.0 and 55°C Streptomyces griseus

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.10.3.4 23000
-
 Superdex 200 gel filtration Streptomyces griseus
1.10.3.15 36000
-
 x * 36000, SDS-PAGE Streptomyces griseus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.10.3.15 3-amino-4-hydroxybenzoate + N-acetyl-L-cysteine + O2 Streptomyces griseus
-
 grixazone B + H2O + CO2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.10.3.4 Streptomyces griseus
-
-
-
1.10.3.15 Streptomyces griseus
-
-
-
1.10.3.15 Streptomyces griseus IFO 13350
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.10.3.4 Cu2+-charged HiTrap chelating HP column chromatography, HiTrap Q column chromatography and Resource PHE column chromatography Streptomyces griseus
1.10.3.15 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]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.10.3.4 0.0000035
-
 crude cell extract, at 30°C Streptomyces griseus
1.10.3.4 0.066
-
 after 19000fold purification, at 30°C Streptomyces griseus
1.10.3.15 0.0000035
-
 crude extract, at pH 7.0 and 30°C Streptomyces griseus
1.10.3.15 0.0658
-
 after 19000fold purification, at pH 7.0 and 30°C Streptomyces griseus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.10.3.4 2-amino-4-methylphenol + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.4 2-amino-4-methylphenol + O2
-
 Streptomyces griseus IFO 13350 ?
-
 ?
1.10.3.4 3,4-dihydroxybenzaldehyde + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.4 3,4-dihydroxybenzaldehyde + O2
-
 Streptomyces griseus IFO 13350 ?
-
 ?
1.10.3.4 3-amino-4-hydroxybenzaldehyde + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.4 catechol + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.4 catechol + O2
-
 Streptomyces griseus IFO 13350 ?
-
 ?
1.10.3.4 L-DOPA + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.4 L-DOPA + O2
-
 Streptomyces griseus IFO 13350 ?
-
 ?
1.10.3.4 o-aminophenol + O2
-
 Streptomyces griseus 2-aminophenoxazin-3-one + H2O
-
 ?
1.10.3.4 protocatechuic acid + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.4 protocatechuic acid + O2
-
 Streptomyces griseus IFO 13350 ?
-
 ?
1.10.3.15 2-amino-4-methylphenol + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.15 3,4-dihydroxybenzaldehyde + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.15 3-amino-4-hydroxybenzaldehyde + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus grixazone A + H2O + CO2
-
 ?
1.10.3.15 3-amino-4-hydroxybenzoate + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus grixazone B + H2O + CO2
-
 ?
1.10.3.15 4-amino-3-hydroxybenzoate + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.15 catechol + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.15 L-Dopa + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus ?
-
 ?
1.10.3.15 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 ?
-
 ?
1.10.3.15 o-aminophenol + N-acetyl-L-cysteine + O2
-
 Streptomyces griseus 2-aminophenoxazin-3-one + H2O + CO2
-
 ?

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
1.10.3.4 GriF
-
 Streptomyces griseus
1.10.3.15 GriF
-
 Streptomyces griseus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.10.3.4 55
-
 at pH 7.0 Streptomyces griseus
1.10.3.15 55
-
 at pH 7.0 Streptomyces griseus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.10.3.15 40 70 more than 50% activity between 30 and 70°C Streptomyces griseus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.10.3.4 55
-
 the enzyme remains active below 55°C at pH 7.0 for 20 min at various temperatures Streptomyces griseus
1.10.3.15 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.10.3.4 0.066
-
 L-Dopa in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 0.8
-
 3,4-Dihydroxybenzaldehyde in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 12
-
 catechol in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 14
-
 3-amino-4-hydroxybenzaldehyde in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 18
-
 2-amino-4-methylphenol in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.4 20
-
 o-aminophenol in 50 mM sodium phosphate, pH 7.0, 30°C Streptomyces griseus
1.10.3.15 0.066
-
 L-Dopa at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 0.8
-
 3,4-Dihydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 12
-
 catechol at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 14
-
 3-amino-4-hydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 18
-
 2-amino-4-methylphenol at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 20
-
 o-aminophenol at pH 7.0 and 55°C Streptomyces griseus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.10.3.4 8.5 10.5
-
 Streptomyces griseus
1.10.3.15 8.5 10.5
-
 Streptomyces griseus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.10.3.4 6 11
-
 Streptomyces griseus
1.10.3.15 6 11 more than 50% activity between pH 6.0 and 11.0 Streptomyces griseus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.10.3.15 6.5 8.5 the enzyme is most stable at pH 6.5-8.5 Streptomyces griseus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.10.3.4 1.9
-
 p-Hydroxybenzaldehyde
-
 Streptomyces griseus
1.10.3.4 3.2
-
 L-tyrosine
-
 Streptomyces griseus
1.10.3.4 3.9
-
 4-hydroxy-3-nitrobenzaldehyde
-
 Streptomyces griseus
1.10.3.4 6.1
-
 o-Nitrophenol
-
 Streptomyces griseus
1.10.3.4 14
-
 aniline
-
 Streptomyces griseus
1.10.3.4 21
-
 phenol
-
 Streptomyces griseus
1.10.3.4 31
-
 3-amino-4-hydroxybenzensulfonic acid
-
 Streptomyces griseus
1.10.3.15 1.9
-
 4-hydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 3.2
-
 L-tyrosine at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 3.9
-
 4-hydroxy-3-nitrobenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 6.1
-
 2-Nitrophenol at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 14
-
 aniline at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 21
-
 phenol at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 31
-
 3-amino-4-hydroxybenzensulfonic acid at pH 7.0 and 55°C Streptomyces griseus

General Information

EC Number General Information Comment Organism
1.10.3.15 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.10.3.15 0.0073
-
 4-Amino-3-hydroxybenzoate at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 0.012
-
 L-Dopa at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 0.19
-
 3-amino-4-hydroxybenzoate at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 0.32
-
 catechol at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 0.97
-
 3,4-Dihydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 5.8
-
 o-aminophenol at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 23
-
 2-amino-4-methylphenol at pH 7.0 and 55°C Streptomyces griseus
1.10.3.15 24
-
 3-amino-4-hydroxybenzaldehyde at pH 7.0 and 55°C Streptomyces griseus