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Literature summary for 1.11.1.16 extracted from

  • Vignali, E.; Tonin, F.; Pollegioni, L.; Rosini, E.
    Characterization and use of a bacterial lignin peroxidase with an improved manganese-oxidative activity (2018), Appl. Microbiol. Biotechnol., 102, 10579-10588 .
    View publication on PubMed
Search for more literature for 1.11.1.16

Application

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(Commentary)

Cloned (Comment) Organism
design of a synthetic gene encoding the N246Avariant 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

Protein Variants Comment Organism
N246A site-directed mutagenesis, mutant Rh_DypB Rhodococcus jostii

Inhibitors

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 [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

Metals/Ions Comment Organism Structure
Mn2+ activates Rhodococcus jostii
NaCl activates 2 fold at 1 M, pH 5.0, 25°C Rhodococcus jostii

Organism

Organism UniProt Comment Textmining
Rhodococcus jostii Q0SE24
-
-

Purification (Commentary)

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 and Products (Substrate)

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

Subunits Comment Organism
More determination of the secondary structure of the N246Avariant with alpha-helix and beta-strand content of about 29% and about 19%, respectively Rhodococcus jostii

Synonyms

Synonyms Comment Organism
bacterial lignin peroxidase
-
 Rhodococcus jostii
DypB
-
 Rhodococcus jostii
More see also EC 1.11.1.14, cf. EC 1.11.1.19 Rhodococcus jostii

Temperature Optimum [°C]

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 Range [°C]

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 [°C]

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 [1/s]

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

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 pH Stability Maximum Comment Organism
6 7 recombinant enzyme, stable at Rhodococcus jostii

kcat/KM [mM/s]

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