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Literature summary for 1.11.1.13 extracted from

  • Li, L.; Liu, B.; Yang, J.; Zhang, Q.; He, C.; Jia, R.
    Catalytic properties of a short manganese peroxidase from Irpex lacteus F17 and the role of Glu166 in the Mn2+-independent activity (2019), Int. J. Biol. Macromol., 136, 859-869 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene ilmnp1, recombinant expression of isozyme Il-MnP1 Irpex lacteus

Protein Variants

Protein Variants Comment Organism
E166D site-directed mutagenesis, the E166D mutant shows no obvious improvement to Mn2+-independent oxidative activity of Il-MnP1 Irpex lacteus
E166G site-directed mutagenesis, the mutant shows highly improved Mn2+-independent oxidative activity, as compared to the wild-type enzyme, with 170fold increased Kcat/Km value. Mutant E166G exhibits 27, 17, 75, 14, and 29fold increase to Mn2+-independent oxidative activity of Il-MnP1 for the phenolic substrates (DMP, guaiacol, catechol, HQ) and the nonphenolic substrate (ABTS), respectively, compared to wild-type Irpex lacteus
E166Q site-directed mutagenesis, the mutant shows highly improved Mn2+-independent oxidative activity, as compared to the wild-type enzyme, with 34fold increased Kcat/Km value. The E166Q mutant displays a 5fold increase to the oxidative activity of Il-MnP1 for all the substrates compared to wild-type Il-MnP1 Irpex lacteus
additional information both wild-type Il-MnP1 and the variants exhibit negligible activity on veratryl alcohol oxidation in the absence of Mn2+ Irpex lacteus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information steady-state kinetics Irpex lacteus
0.0156
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
0.0365
-
guaiacol pH 4.0, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
0.096
-
2,6-dimethoxyphenol pH 7.4, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
0.195
-
guaiacol pH 7.4, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
0.271
-
guaiacol pH 7.4, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
0.435
-
2,6-dimethoxyphenol pH 7.4, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
1.3
-
guaiacol pH 4.0, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
3.03
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant mutant E166G, in absence of Mn2+ Irpex lacteus
3.76
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant mutant E166Q, in absence of Mn2+ Irpex lacteus
3.96
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant mutant E166D, in absence of Mn2+ Irpex lacteus
4.63
-
guaiacol pH 4.0, 30°C, recombinant mutant E166Q, in absence of Mn2+ Irpex lacteus
5.75
-
guaiacol pH 4.0, 30°C, recombinant mutant E166G, in absence of Mn2+ Irpex lacteus
6.31
-
guaiacol pH 4.0, 30°C, recombinant mutant E166D, in absence of Mn2+ Irpex lacteus
7.23
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ Mn2+ binds to MnPs in a site that is composed of three acidic amino acids. Isozyme Il-MnP1 exhibits higher oxidative activity in the presence of Mn2+ than in the absence of Mn2+ toward the majority of the selected substrates at pH 4.0 Irpex lacteus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 Mn(II) + 2 H+ + H2O2 Irpex lacteus
-
2 Mn(III) + 2 H2O
-
?
2 Mn(II) + 2 H+ + H2O2 Irpex lacteus F17
-
2 Mn(III) + 2 H2O
-
?
2 Mn(II) + 2 H+ + H2O2 Irpex lacteus CCTCC AF2014020
-
2 Mn(III) + 2 H2O
-
?

Organism

Organism UniProt Comment Textmining
Irpex lacteus S4W784
-
-
Irpex lacteus CCTCC AF2014020 S4W784
-
-
Irpex lacteus F17 S4W784
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
oxidative activity for phenols/non-phenols by recombinant isozyme Il-MnP1 in the presence/absence of Mn2+ in U/l, overview. For monophenols (DMP, guaiacol) without Mn2+, the reaction results in a 95% reduction of oxidative activity, whereas bisphenols are only found to obtain a 20% reduction (catechol) or 50% increase (HQ). As for non-phenols, veratryl alcohol is oxidized only when Mn2+ is present, while Il-MnP1 still retains 70% of its oxidative activity toward ABTS in the absence of Mn2+ Irpex lacteus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,4-benzenediol + 2 H+ + H2O2
-
Irpex lacteus ? + 2 H2O
-
?
1,4-benzenediol + 2 H+ + H2O2
-
Irpex lacteus F17 ? + 2 H2O
-
?
1,4-benzenediol + 2 H+ + H2O2
-
Irpex lacteus CCTCC AF2014020 ? + 2 H2O
-
?
2 Mn(II) + 2 H+ + H2O2
-
Irpex lacteus 2 Mn(III) + 2 H2O
-
?
2 Mn(II) + 2 H+ + H2O2
-
Irpex lacteus F17 2 Mn(III) + 2 H2O
-
?
2 Mn(II) + 2 H+ + H2O2
-
Irpex lacteus CCTCC AF2014020 2 Mn(III) + 2 H2O
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H+ + H2O2 ABTS Irpex lacteus oxidized 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H2O
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H+ + H2O2 ABTS Irpex lacteus F17 oxidized 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H2O
-
?
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H+ + H2O2 ABTS Irpex lacteus CCTCC AF2014020 oxidized 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H2O
-
?
2,6-dimethoxyphenol + 2 H+ + H2O2
-
Irpex lacteus oxidized 2,6-dimethoxyphenol + 2 H2O
-
?
2,6-dimethoxyphenol + 2 H+ + H2O2
-
Irpex lacteus F17 oxidized 2,6-dimethoxyphenol + 2 H2O
-
?
2,6-dimethoxyphenol + 2 H+ + H2O2
-
Irpex lacteus CCTCC AF2014020 oxidized 2,6-dimethoxyphenol + 2 H2O
-
?
catechol + 2 H+ + H2O2 1,2-benzenediol Irpex lacteus ? + 2 H2O
-
?
guaiacol + 2 H+ + H2O2
-
Irpex lacteus oxidized guaiacol + 2 H2O
-
?
guaiacol + 2 H+ + H2O2 Il-MnP1 oxidizes guaiacol and provides a first radical A, which undergoes a variety of non-enzymatic reactions that mainly consists of reactions of resonance stabilization to generate the next radical B. In turn the C-C radical coupling of two B radicals generates a dimeric 3',5-dimethoxy-3,4-dihydro[1,1'-biphenyl]-4,4'-diol, which can be further oxidized by Il-MnP1 to produce 3,3'-dimethoxy[1,1'-bi(cyclohexa-2,5-diene)]-4,4'-dione and 1-[[1(1')Z]-3'-methoxy-4,4'-dioxo[1,1'-bi(cyclohexa-2,5-dien-1-yliden)]-3-yl]-1-methyldioxidan-1-ium. Furthermore, Il-MnP1 oxidizes 3',5-dimethoxy-3,4-dihydro[1,1'-biphenyl]-4,4'-diol to produce a radical C,which is also subject to a variety of non-enzymatic reactions to produce a radical D. Finally, radical D is subjected to further oxidation and C-C coupling for the production of 4'-hydroxy-3',5-dimethoxy[1,1'-biphenyl]-3,4-dione, 1(3),2(5),3(3)-trimethoxy-2(3),2(4)-dihydro[1(1),2(1):2(3),3(1)-terphenyl]-1(4),2(4),3(4)-triol, and [1(1)(2(1))E]-3(4)-hydroxy-1(3),2(5),3(3)-trimethoxy-1(4)H,2(4)H-[1(1),2(1):2(3),3(1)-terphenyl]-1(4),2(4)-dione Irpex lacteus oxidized guaiacol 4'-hydroxy-3',5-dimethoxy[1,1'-biphenyl]-3,4-dione + 2 H2O
-
?
additional information the oxidation of guaiacol mainly belongs to a series of polymeric reactions of radicals initiated by isozyme Il-MnP1,whether they are in the presence and absence of Mn2+ at either pH 4.0 or pH 7.4. Both wild-type Il-MnP1 and the variants exhibit negligible activity on veratryl alcohol oxidation in the absence of Mn2+ Irpex lacteus ?
-
-
additional information the oxidation of guaiacol mainly belongs to a series of polymeric reactions of radicals initiated by isozyme Il-MnP1,whether they are in the presence and absence of Mn2+ at either pH 4.0 or pH 7.4. Both wild-type Il-MnP1 and the variants exhibit negligible activity on veratryl alcohol oxidation in the absence of Mn2+ Irpex lacteus F17 ?
-
-
additional information the oxidation of guaiacol mainly belongs to a series of polymeric reactions of radicals initiated by isozyme Il-MnP1,whether they are in the presence and absence of Mn2+ at either pH 4.0 or pH 7.4. Both wild-type Il-MnP1 and the variants exhibit negligible activity on veratryl alcohol oxidation in the absence of Mn2+ Irpex lacteus CCTCC AF2014020 ?
-
-
veratryl alcohol + H2O2
-
Irpex lacteus 3,4-dimethoxybenzoic acid + 2 H2O
-
?

Synonyms

Synonyms Comment Organism
Il-MnP1
-
Irpex lacteus
MnP
-
Irpex lacteus
short manganese peroxidase
-
Irpex lacteus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Irpex lacteus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.75
-
guaiacol pH 7.4, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
1.82
-
2,6-dimethoxyphenol pH 7.4, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
11.26
-
guaiacol pH 4.0, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
20.54
-
guaiacol pH 7.4, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
26.15
-
guaiacol pH 4.0, 30°C, recombinant mutant E166D, in absence of Mn2+ Irpex lacteus
37.86
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
64.57
-
guaiacol pH 4.0, 30°C, recombinant mutant E166Q, in absence of Mn2+ Irpex lacteus
80.1
-
2,6-dimethoxyphenol pH 7.4, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
136
-
guaiacol pH 4.0, 30°C, recombinant mutant E166G, in absence of Mn2+ Irpex lacteus
142
-
guaiacol pH 4.0, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
148
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
379
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant mutant E166D, in absence of Mn2+ Irpex lacteus
647
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant mutant E166Q, in absence of Mn2+ Irpex lacteus
2590
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant mutant E166G, in absence of Mn2+ Irpex lacteus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
when comparing the oxidative activity of Il-MnP1 between pH 4.0 and pH 7.4, the oxidative activity of Il-MnP1 at pH 4.0 is usually higher than that at pH 7.4. But there are some exceptions Irpex lacteus
4
-
peak 1 Irpex lacteus
7.4
-
peak 2 Irpex lacteus

Cofactor

Cofactor Comment Organism Structure
heme
-
Irpex lacteus

General Information

General Information Comment Organism
evolution the enzyme belongs to the class II fungal peroxidases (PODs) Irpex lacteus
additional information role of Glu166 in the Mn2+-independent activity Irpex lacteus
physiological function class II fungal peroxidases (PODs), manganese peroxidases (MnPs) are named for their strongMn2+-dependent oxidative activity for an array of aromatic substrates, including phenols, nonphenols, and various industrial dyes Irpex lacteus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.85
-
guaiacol pH 7.4, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
4.14
-
guaiacol pH 4.0, 30°C, recombinant mutant E166D, in absence of Mn2+ Irpex lacteus
4.18
-
2,6-dimethoxyphenol pH 7.4, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
5.24
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
8.66
-
guaiacol pH 4.0, 30°C, recombinant wild-type enzyme, in absence of Mn2+ Irpex lacteus
13.95
-
guaiacol pH 4.0, 30°C, recombinant mutant E166Q, in absence of Mn2+ Irpex lacteus
72
-
guaiacol pH 4.0, 30°C, recombinant mutant E166G, in absence of Mn2+ Irpex lacteus
75.79
-
guaiacol pH 7.4, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
95.7
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant mutant E166D, in absence of Mn2+ Irpex lacteus
172.1
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant mutant E166Q, in absence of Mn2+ Irpex lacteus
834.4
-
2,6-dimethoxyphenol pH 7.4, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
854.8
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant mutant E166G, in absence of Mn2+ Irpex lacteus
3890
-
guaiacol pH 4.0, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus
9487
-
2,6-dimethoxyphenol pH 4.0, 30°C, recombinant wild-type enzyme, in presence of Mn2+ Irpex lacteus