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Literature summary extracted from

  • Ogola, H.J.; Kamiike, T.; Hashimoto, N.; Ashida, H.; Ishikawa, T.; Shibata, H.; Sawa, Y.
    Molecular characterization of a novel DyP-type peroxidase from the cyanobacterium Anabaena sp. PCC 7120 (2009), Appl. Environ. Microbiol., 75, 7509-7518.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.11.1.7 syringaldehyde the decolorization activity of AnaPX toward azo dyes is significantly enhanced 2-50fold in the presence of syringaldehyde Anabaena sp.
1.11.1.19 syringaldehyde the presence of syringaldehyde drastically enhances the rate of decolorization of Reactive Black 5, Reactive Orange 14, Reactive Red 120, Reactive Green 19, and Acid Red with 50fold, 9fold, 15fold, 2fold, and 7fold improvements, respectively Anabaena sp.

Application

EC Number Application Comment Organism
1.11.1.19 additional information AnaPX is a useful alternative of horseradish peroxidase or fungal DyPs Anabaena sp.

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.11.1.7 expressed in Escherichia coli BL21(DE3) cells Anabaena sp.
1.11.1.19 expressed in Escherichia coli BL21(DE3) cells Anabaena sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.11.1.7 diethyl dicarbonate less than 50% inhibition at 50 mM Anabaena sp.
1.11.1.7 Fe2+ 98.8% inhibition at 5 mM Anabaena sp.
1.11.1.7 KCN very low sensitivity, causes only 9% and 14% inhibition of the activity at 1 and 10 mM, respectively Anabaena sp.
1.11.1.7 Mn2+ 57.1% inhibition at 5 mM Anabaena sp.
1.11.1.7 additional information the enzyme is not sensitive to the prototypical catalase inhibitor 3-amino-1, 2, 4,-triazole in presence of ascorbic acid (1.0 mM) Anabaena sp.
1.11.1.7 NaN3 82% inhibition 10 mM Anabaena sp.
1.11.1.7 phenylhydrazine complete inhibition at 1 mM Anabaena sp.
1.11.1.19 diethyl dicarbonate less than 50% inhibition at 50 mM Anabaena sp.
1.11.1.19 Fe2+ AnaPX is highly sensitive to Fe2+ (98.8% inhibition at 5 mM) Anabaena sp.
1.11.1.19 KCN AnaPX shows very low sensitivity to KCN which causes only 9% and 14% inhibition of the activity at 1 and 10 mM, respectively Anabaena sp.
1.11.1.19 Mn2+ 57.1% inhibition at 5 mM Anabaena sp.
1.11.1.19 additional information the enzyme is not sensitive to the prototypical catalase inhibitor 3-amino-1,2,4,-triazole in the presence of ascorbic acid (1.0 mM), metal chelating and sulfhydryl reagents do not significantly affect activity Anabaena sp.
1.11.1.19 NaN3 AnaPX is highly sensitive to NaN3 (82% inhibition at 10 mM) Anabaena sp.
1.11.1.19 phenylhydrazine AnaPX is highly sensitive to the suicide substrate phenylhydrazine (complete inhibition at 1 mM) Anabaena sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.11.1.7 0.0036
-
 Reactive Blue 5 at pH 4.8 Anabaena sp.
1.11.1.7 0.0058
-
 H2O2 at pH 4.8 Anabaena sp.
1.11.1.19 0.0036
-
 Reactive Blue 5 in 50 mM citrate buffer (pH 4.0-4.4), at 37°C Anabaena sp.
1.11.1.19 0.0058
-
 H2O2 in 50 mM citrate buffer (pH 4.0-4.4), at 37°C Anabaena sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.11.1.7 53280
-
 4 * 53280, MALDI-TOF mass spectrometry Anabaena sp.
1.11.1.7 53368
-
 4 * 53368, calculated from amino acid sequence Anabaena sp.
1.11.1.7 54000
-
 4 * 54000, SDS-PAGE Anabaena sp.
1.11.1.19 53280
-
 4 * 53280, native enzyme, MALDI-TOF spectrometry Anabaena sp.
1.11.1.19 53368
-
 4 * 53368, deduced from amino acid sequence Anabaena sp.
1.11.1.19 54000
-
 4 * 54000, native enzyme, SDS-PAGE Anabaena sp.
1.11.1.19 209000
-
 native enzyme, gel filtration Anabaena sp.

Organism

EC Number Organism UniProt Comment Textmining
1.11.1.7 Anabaena sp.
-
 strain PCC 7120
-
1.11.1.19 Anabaena sp. Q8YWM0 strain PCC7120 (ATCC 27893)
-

Oxidation Stability

EC Number Oxidation Stability Organism
1.11.1.7 retains more than 50% of its activity at 2.5 mM H2O2 Anabaena sp.
1.11.1.19 the enzyme retains more than 50% of its activity at 2.5 mM H2O2 Anabaena sp.

Purification (Commentary)

EC Number Purification (Comment) Organism
1.11.1.7 DEAE-Toyopearl column chromatography, ammonium sulfate precipitation, butyl-Toyopearl column chromatography, and hydroxylapatite column chromatography Anabaena sp.
1.11.1.19 Toyopearl DEAE-650M column chromatography, Toyopearl butyl-650M column chromatography, and hydroxylapatite column chromatography Anabaena sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.11.1.7 5.6
-
 recombinant enzyme from crude extract Anabaena sp.
1.11.1.7 283
-
 recombinant enzyme after 50.5fold purification Anabaena sp.
1.11.1.19 5.6
-
 recombinant enzyme from crude extract, using Reactive Blue 5 as substrate, pH 4.0-4.4, at 37°C Anabaena sp.
1.11.1.19 54
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using syringaldehyde as substrate Anabaena sp.
1.11.1.19 71
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using 2,6-dimethoxyphenol as substrate Anabaena sp.
1.11.1.19 204
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using 2,2'-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid as substrate Anabaena sp.
1.11.1.19 216
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using pyrogallol as substrate Anabaena sp.
1.11.1.19 230
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using guaiacol as substrate Anabaena sp.
1.11.1.19 283
-
 recombinant enzyme after 50.5fold purification, using Reactive Blue 5 as substrate, pH 4.0-4.4, at 37°C Anabaena sp.
1.11.1.19 290
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using L-ascorbate as substrate Anabaena sp.
1.11.1.19 308
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using D-isoascorbate as substrate Anabaena sp.
1.11.1.19 308
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using NADPH as substrate Anabaena sp.
1.11.1.19 374
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using NADH as substrate Anabaena sp.
1.11.1.19 1478
-
 recombinant enzyme, in 50 mM citrate buffer (pH 4.0-4.4), at 37°C, using 4-aminoantipyrine as substrate Anabaena sp.

Storage Stability

EC Number Storage Stability Organism
1.11.1.7 4°C, in 50 mM potassium phosphate (pH 7.0), 40 days, less than 10% loss of activity Anabaena sp.
1.11.1.19 4°C, 50 mM potassium phosphate buffer (pH 7.0), 40 days, 10% loss of activity Anabaena sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.11.1.7 2,6-dimethoxyphenol + H2O2
-
 Anabaena sp. ?
-
 ?
1.11.1.7 4-aminoantipyrine + H2O2 643% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.7 ABTS + H2O2 89% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.7 Acid Blue 45 + H2O2
-
 Anabaena sp. ?
-
 ?
1.11.1.7 D-iso-ascorbate + H2O2 134% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.7 guaiacol + H2O2 100% activity Anabaena sp. ?
-
 ?
1.11.1.7 L-ascorbate + H2O2 126% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.7 additional information no activity with Mn2+ and veratryl alcohol as electron donors Anabaena sp. ?
-
 ?
1.11.1.7 NADH + H2O2 163% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.7 NADPH + H2O2 134% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.7 pyrogallol + H2O2 93% activity compared to guaiacol Anabaena sp. purpurogallin + H2O
-
 ?
1.11.1.7 Reactive Blue 114 + H2O2
-
 Anabaena sp. ?
-
 ?
1.11.1.7 Reactive Blue 119 + H2O2
-
 Anabaena sp. ?
-
 ?
1.11.1.7 Reactive Blue 4 + H2O2
-
 Anabaena sp. ?
-
 ?
1.11.1.7 Reactive Blue 5 + H2O2 123% activity compared to guaiacol, the apparent optimum concentration of H2O2 required for the decolorization of Reactive Blue 5 by AnaPX is 0.4 mM Anabaena sp. ?
-
 ?
1.11.1.7 syringaldehyde + H2O2 23% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.19 2,2'-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid + H2O2 + H+ 89% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.19 2,6-dimethoxyphenol + H2O2 + H+ 31% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.19 4-aminoantipyrine + H2O2 + H+ 643% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.19 Acid Blue 45 + H2O2 + H+ 98% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Acid Blue 45 + H2O
-
 ?
1.11.1.19 Acid Red 151 + H2O2 + H+ 12% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Acid Red 151 + H2O
-
 ?
1.11.1.19 Congo Red + H2O2 + H+ 12% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Congo Red + H2O
-
 ?
1.11.1.19 D-isoascorbate + H2O2 + H+ 134% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.19 Direct Sky Blue 6B + H2O2 + H+ 50% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Direct Sky Blue 6B + H2O
-
 ?
1.11.1.19 guaiacol + H2O2 + H+ 100% activity Anabaena sp. ?
-
 ?
1.11.1.19 L-ascorbate + H2O2 + H+ 126% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.19 additional information the decolorization activity of AnaPX toward azo dyes is relatively low. No decolorization of Toluidine Blue O, Reactive Red 33, and Reactive Yellow 2 is observed Anabaena sp. ?
-
 ?
1.11.1.19 Procion Blue H-ERD + H2O2 + H+ 90% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Procion Blue H-ERD + H2O
-
 ?
1.11.1.19 Procion Blue H-EXL + H2O2 + H+ 72% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Procion Blue H-EXL + H2O
-
 ?
1.11.1.19 pyrogallol + H2O2 + H+ 93% activity compared to guaiacol Anabaena sp. ?
-
 ?
1.11.1.19 Reactive Black 5 + H2O2 + H+ 8% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Reactive Black 5 + H2O
-
 ?
1.11.1.19 Reactive Blue 114 + H2O2 + H+ 187% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Reactive Blue 114 + H2O
-
 ?
1.11.1.19 Reactive Blue 19 + H2O2 + H+ 153% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Reactive Blue 19 + H2O
-
 ?
1.11.1.19 Reactive Blue 4 + H2O2 + H+ 64% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Reactive Blue 4 + H2O
-
 ?
1.11.1.19 Reactive Blue 5 + H2O2 + H+ 123% activity compared to guaiacol, the apparent optimum concentration of H2O2 required for the decolorization of Reactive Blue 5 by AnaPX is 0.4 mM Anabaena sp. oxidized Reactive Blue 5 + H2O
-
 ?
1.11.1.19 Reactive Green 19 + H2O2 + H+ 35% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Reactive Green 19 + H2O
-
 ?
1.11.1.19 Reactive Orange 14 + H2O2 + H+ 10% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Reactive Orange 14 + H2O
-
 ?
1.11.1.19 Reactive Red 120 + H2O2 + H+ 5% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Reactive Red 120 + H2O
-
 ?
1.11.1.19 Reactive Yellow 86 + H2O2 + H+ 3% decolorization rate compared to Reactive Blue 5 Anabaena sp. oxidized Reactive Yellow 86 + H2O
-
 ?
1.11.1.19 syringaldehyde + H2O2 + H+ 23% activity compared to guaiacol Anabaena sp. oxidized syringaldehyde + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.11.1.7 homotetramer 4 * 54000, SDS-PAGE Anabaena sp.
1.11.1.7 homotetramer 4 * 53280, MALDI-TOF mass spectrometry Anabaena sp.
1.11.1.7 homotetramer 4 * 53368, calculated from amino acid sequence Anabaena sp.
1.11.1.19 homotetramer 4 * 53280, native enzyme, MALDI-TOF spectrometry Anabaena sp.
1.11.1.19 homotetramer 4 * 53368, deduced from amino acid sequence Anabaena sp.
1.11.1.19 homotetramer 4 * 54000, native enzyme, SDS-PAGE Anabaena sp.

Synonyms

EC Number Synonyms Comment Organism
1.11.1.7 AnaPX
-
 Anabaena sp.
1.11.1.19 AnaPX
-
 Anabaena sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.11.1.7 35
-
-
 Anabaena sp.
1.11.1.19 35
-
-
 Anabaena sp.

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.11.1.7 30 60 the enzyme is considerably more stable at 30°C and 40°C where it retains more than 90% of its activity, the enzyme loses more than 90% of its activity after incubation at 50°C and 60°C for 3 h Anabaena sp.
1.11.1.19 30 60 the enzyme is considerably stable at 30°C and 40°C where it retains more than 90% of its activity, while the enzyme loses more than 90% of its activity after incubation at 50°C and 60°C for 3 h Anabaena sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.11.1.19 384
-
 H2O2 in 50 mM citrate buffer (pH 4.0-4.4), at 37°C Anabaena sp.
1.11.1.19 384
-
 Reactive Blue 5 in 50 mM citrate buffer (pH 4.0-4.4), at 37°C Anabaena sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.11.1.7 4 5
-
 Anabaena sp.
1.11.1.19 4 4.4
-
 Anabaena sp.

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.11.1.7 3.5 9.5 when maintained at 40°C for 20 min, the enzyme is stable at pH values between 3.5 and 9.5 Anabaena sp.
1.11.1.19 3.5 9.5 when maintained at 40°C for 20 min, the enzyme is stable at pH values between 3.5 and 9.5. The enzyme shows 20% relative activity at pH 3.0 and 10.0, 40% relative activity at pH 4.0, about 65% relative activity at pH 4.5 and 9.0, about 80% relative activity at pH 5.0, more than 90% relative activity at pH 6.0-8.0 Anabaena sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.11.1.7 heme iron protoporphyrin-containing heme peroxidase Anabaena sp.
1.11.1.19 heme iron protoporphyrin-containing heme peroxidase, the heme content is estimated to be 0.91 mol per mole of protein Anabaena sp.

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.11.1.7 Anabaena sp. isoelectric focusing
-
 3.7
1.11.1.19 Anabaena sp. isoelectric focusing
-
 3.7

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.11.1.19 66000
-
 H2O2 in 50 mM citrate buffer (pH 4.0-4.4), at 37°C Anabaena sp.
1.11.1.19 120000
-
 Reactive Blue 5 in 50 mM citrate buffer (pH 4.0-4.4), at 37°C Anabaena sp.