Application | Comment | Organism |
---|---|---|
additional information | peroxidases are well-known biocatalysts produced by all organisms, especially microorganisms, and used in a number of biotechnological applications | Rhodococcus jostii |
Cloned (Comment) | Organism |
---|---|
design of a synthetic gene encoding the N246A variant of DypB from Rhodococcus jostii strain RHA1 (Rh_DypB) is designed by back translation of the protein sequence, recombinant overexpression of His-tagged N246A mutant enzyme in Escherichia coli strain BL21(DE3), the enzyme is fully produced as folded holoenzyme, thus without the need for a further reconstitution step | Rhodococcus jostii |
Protein Variants | Comment | Organism |
---|---|---|
N246A | site-directed mutagenesis, mutant Rh_DypB | Rhodococcus jostii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
DMSO | 50% inhibition at 20%, 90% inhibition at 30%, pH 5.0, 25°C | Rhodococcus jostii | |
Tween-80 | 80% inhibition at 5%, pH 5.0, 25°C | Rhodococcus jostii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics by mutant enzyme N246A | Rhodococcus jostii | |
0.03 | - |
H2O2 | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 50 mM 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | Rhodococcus jostii | |
0.04 | - |
H2O2 | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 50 mM 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2 mM Mn2+ | Rhodococcus jostii | |
3.1 | - |
Mn(II) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 0.3 mM H2O2 | Rhodococcus jostii | |
11.2 | - |
Mn(II) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 1.5 mM H2O2 | Rhodococcus jostii | |
14.9 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 1.5 mM H2O2 | Rhodococcus jostii | |
15.6 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 0.3 mM H2O2 | Rhodococcus jostii | |
17.5 | - |
2,6-dimethoxyphenol | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 0.3 mM H2O2 | Rhodococcus jostii | |
29.5 | - |
2,6-dimethoxyphenol | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 1.5 mM H2O2 | Rhodococcus jostii |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | activates | Rhodococcus jostii | |
NaCl | activates 2 fold at 1 M, pH 5.0, 25°C | Rhodococcus jostii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus jostii | Q0SE24 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged N246A mutant enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Rhodococcus jostii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-phenyl-1,2-ethandiol + H2O2 | substrate of N246A mutant enzyme | Rhodococcus jostii | ? + 2 H2O | - |
? | |
1-phenyl-1-propanol + H2O2 | substrate of N246A mutant enzyme | Rhodococcus jostii | ? + 2 H2O | - |
? | |
1-phenyl-2-propanol + H2O2 | substrate of N246A mutant enzyme | Rhodococcus jostii | ? + 2 H2O | - |
? | |
1-phenylethanol + H2O2 | substrate of N246A mutant enzyme | Rhodococcus jostii | ? + 2 H2O | - |
? | |
2 Mn(II) + 2 H+ + H2O2 | activity of EC 1.11.1.13 | Rhodococcus jostii | 2 Mn(III) + 2 H2O | - |
? | |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H+ + H2O2 | - |
Rhodococcus jostii | oxidized 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) + 2 H2O | - |
? | |
2,6-dimethoxyphenol + 2 H+ + H2O2 | - |
Rhodococcus jostii | oxidized 2,6-dimethoxyphenol + 2 H2O | - |
? | |
Azure B + 2 H+ + H2O2 | dye decolorization, substrate of N246A mutant enzyme | Rhodococcus jostii | oxidized Azure B + 2 H2O | - |
? | |
guaiacylglycerol-beta-guaiacyl ether + H2O2 | GGE, substrate of mutant enzyme N246A | Rhodococcus jostii | glycerol + guaiacol + 2 H2O | - |
? | |
additional information | enzyme DypB degrades solvent-obtained fractions of a Kraft lignin. The recombinant mutant enzyme Rh_DypB shows a classical peroxidase activity which is significantly increased by adding Mn2+ ions, kinetic parameters for H2O2, Mn2+, ABTS, and 2,6-DMP are determined. The enzyme shows broad dye-decolorization activity, especially in the presence of Mn2+, oxidizes various aromatic monomers from lignin, and cleaves the guaiacylglycerol-beta-guaiacyl ether (GGE), i.e., the Calpha-Cbeta bond of the dimeric lignin model molecule of beta-O-4 linkages. Under optimized conditions, 2 mM GGE is fully cleaved by recombinant Rh_DypB, generating guaiacol in only 10 min, at a rate of 12.5 micromol/min/mg enzyme. Screening of oxidation activity on monomeric lignin model compounds, overview | Rhodococcus jostii | ? | - |
- |
|
Reactive Black 5 + 2 H+ + H2O2 | dye decolorization, substrate of N246A mutant enzyme | Rhodococcus jostii | oxidized Reactive Black 5 + 2 H2O | - |
? | |
remazol brilliant blue R + H2O2 | substrate of N246A mutant enzyme | Rhodococcus jostii | oxidized remazol brilliant blue R + 2 H2O | - |
? | |
veratryl alcohol + H2O2 | substrate of N246A mutant enzyme | Rhodococcus jostii | 3,4-dimethoxybenzoic acid + 2 H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | determination of the secondary structure of the N246A variant with alpha-helix and beta-strand content of about 29% and about 19%, respectively | Rhodococcus jostii |
Synonyms | Comment | Organism |
---|---|---|
bacterial lignin peroxidase | - |
Rhodococcus jostii |
DypB | - |
Rhodococcus jostii |
More | see also EC 1.11.1.16, cf. EC 1.11.1.19 | Rhodococcus jostii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
recombinant enzyme, in absense of Mn2+ | Rhodococcus jostii |
65 | - |
recombinant enzyme, in presense of Mn2+ | Rhodococcus jostii |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
3 | 9 | recombinant enzyme, activity range, in absense of Mn2+ | Rhodococcus jostii |
47 | - |
recombinant enzyme, activity range, in presense of Mn2+ | Rhodococcus jostii |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 37 | the recombinant enzyme maintains a large part of the starting activity following incubation for 24 h in this temperature range | Rhodococcus jostii |
63 | 65 | the recombinant enzyme shows a good thermostability with a melting temperature of 63-65°C | Rhodococcus jostii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.45 | - |
2,6-dimethoxyphenol | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 1.5 mM H2O2 | Rhodococcus jostii | |
0.5 | - |
2,6-dimethoxyphenol | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 0.3 mM H2O2 | Rhodococcus jostii | |
12.6 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 1.5 mM H2O2 | Rhodococcus jostii | |
15.4 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 0.3 mM H2O2 | Rhodococcus jostii | |
19 | - |
Mn(II) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 1.5 mM H2O2 | Rhodococcus jostii | |
21.3 | - |
H2O2 | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 50 mM 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | Rhodococcus jostii | |
21.4 | - |
Mn(II) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 0.3 mM H2O2 | Rhodococcus jostii | |
34 | - |
H2O2 | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 50 mM 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2 mM Mn2+ | Rhodococcus jostii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4 | - |
recombinant enzyme, in absense of Mn2+ | Rhodococcus jostii |
6 | - |
recombinant enzyme, in presense of Mn2+ | Rhodococcus jostii |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6 | 7 | recombinant enzyme, stable at | Rhodococcus jostii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
2,6-dimethoxyphenol | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 1.5 mM H2O2 | Rhodococcus jostii | |
0.29 | - |
2,6-dimethoxyphenol | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 0.3 mM H2O2 | Rhodococcus jostii | |
0.85 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 1.5 mM H2O2 | Rhodococcus jostii | |
0.99 | - |
2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 0.3 mM H2O2 | Rhodococcus jostii | |
1.7 | - |
Mn(II) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 1.5 mM H2O2 | Rhodococcus jostii | |
6.9 | - |
Mn(II) | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 0.3 mM H2O2 | Rhodococcus jostii | |
710 | - |
H2O2 | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 50 mM 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) | Rhodococcus jostii | |
850 | - |
H2O2 | pH 5.0, 25°C, recombinant enzyme mutant N246A, in presence of 50 mM 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2 mM Mn2+ | Rhodococcus jostii |