Cloned (Comment) | Organism |
---|---|
gene ilmnp1, recombinant expression of isozyme Il-MnP1 | Irpex lacteus |
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 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 | 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Irpex lacteus | S4W784 | - |
- |
Irpex lacteus CCTCC AF2014020 | S4W784 | - |
- |
Irpex lacteus F17 | S4W784 | - |
- |
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 | 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 | Comment | Organism |
---|---|---|
Il-MnP1 | - |
Irpex lacteus |
MnP | - |
Irpex lacteus |
short manganese peroxidase | - |
Irpex lacteus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Irpex lacteus |
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 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 | Comment | Organism | Structure |
---|---|---|---|
heme | - |
Irpex lacteus |
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 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 |