Formation of a tyrosine adduct involved in lignin degradation by Trametopsis cervina lignin peroxidase: a novel peroxidase activation mechanism

2013 ◽  
Vol 452 (3) ◽  
pp. 575-584 ◽  
Author(s):  
Yuta Miki ◽  
Rebecca Pogni ◽  
Sandra Acebes ◽  
Fátima Lucas ◽  
Elena Fernández-Fueyo ◽  
...  
Keyword(s):  
Redox Potential ◽  
Lignin Peroxidase ◽  
Transient State ◽  
Veratryl Alcohol ◽  
Activation Mechanism ◽  
Rate Limiting Step ◽  
Lignin Model ◽  
Lag Period ◽  
Compound Ii ◽  
Protein Radicals

LiP (lignin peroxidase) from Trametopsis cervina has an exposed catalytic tyrosine residue (Tyr181) instead of the tryptophan conserved in other lignin-degrading peroxidases. Pristine LiP showed a lag period in VA (veratryl alcohol) oxidation. However, VA-LiP (LiP after treatment with H2O2 and VA) lacked this lag, and H2O2-LiP (H2O2-treated LiP) was inactive. MS analyses revealed that VA-LiP includes one VA molecule covalently bound to the side chain of Tyr181, whereas H2O2-LiP contains a hydroxylated Tyr181. No adduct is formed in the Y171N variant. Molecular docking showed that VA binding is favoured by sandwich π stacking with Tyr181 and Phe89. EPR spectroscopy after peroxide activation of the pre-treated LiPs showed protein radicals other than the tyrosine radical found in pristine LiP, which were assigned to a tyrosine–VA adduct radical in VA-LiP and a dihydroxyphenyalanine radical in H2O2-LiP. Both radicals are able to oxidize large low-redox-potential substrates, but H2O2-LiP is unable to oxidize high-redox-potential substrates. Transient-state kinetics showed that the tyrosine–VA adduct strongly promotes (>100-fold) substrate oxidation by compound II, the rate-limiting step in catalysis. The novel activation mechanism is involved in ligninolysis, as demonstrated using lignin model substrates. The present paper is the first report on autocatalytic modification, resulting in functional alteration, among class II peroxidases.

scholarly journals Comparing Ligninolytic Capabilities of Bacterial and Fungal Dye-Decolorizing Peroxidases and Class-II Peroxidase-Catalases

2021 ◽  
Vol 22 (5) ◽  
pp. 2629
Author(s):  
Dolores Linde ◽  
Iván Ayuso-Fernández ◽  
Marcos Laloux ◽  
José E. Aguiar-Cervera ◽  
Antonio L. de Lacey ◽  
...  
Keyword(s):  
Veratryl Alcohol ◽  
Flow Measurements ◽  
Reduction Potentials ◽  
Rate Limiting Step ◽  
Lower Extent ◽  
Lignin Model ◽  
Bacteria And Fungi ◽  
Compound Ii ◽  
Rate Limiting ◽  
Lignin Macromolecule

We aim to clarify the ligninolytic capabilities of dye-decolorizing peroxidases (DyPs) from bacteria and fungi, compared to fungal lignin peroxidase (LiP) and versatile peroxidase (VP). With this purpose, DyPs from Amycolatopsis sp., Thermomonospora curvata, and Auricularia auricula-judae, VP from Pleurotus eryngii, and LiP from Phanerochaete chrysosporium were produced, and their kinetic constants and reduction potentials determined. Sharp differences were found in the oxidation of nonphenolic simple (veratryl alcohol, VA) and dimeric (veratrylglycerol-β- guaiacyl ether, VGE) lignin model compounds, with LiP showing the highest catalytic efficiencies (around 15 and 200 s−1·mM−1 for VGE and VA, respectively), while the efficiency of the A. auricula-judae DyP was 1–3 orders of magnitude lower, and no activity was detected with the bacterial DyPs. VP and LiP also showed the highest reduction potential (1.28–1.33 V) in the rate-limiting step of the catalytic cycle (i.e., compound-II reduction to resting enzyme), estimated by stopped-flow measurements at the equilibrium, while the T. curvata DyP showed the lowest value (1.23 V). We conclude that, when using realistic enzyme doses, only fungal LiP and VP, and in much lower extent fungal DyP, oxidize nonphenolic aromatics and, therefore, have the capability to act on the main moiety of the native lignin macromolecule.

scholarly journals Influence of cellobiose oxidase on peroxidases from Phanerochaete chrysosporium

1993 ◽  
Vol 293 (2) ◽  
pp. 431-435 ◽  
Author(s):  
P Ander ◽  
G Sena-Martins ◽  
J C Duarte
Keyword(s):  
Cytochrome C ◽  
Horseradish Peroxidase ◽  
Phanerochaete Chrysosporium ◽  
Lignin Peroxidase ◽  
Manganese Peroxidase ◽  
Veratryl Alcohol ◽  
Lag Phase ◽  
And Function ◽  
Compound Ii ◽  
Catalytic Cycles

Reduction of H2O2-oxidized manganese peroxidase (MnP), lignin peroxidase and, to some extent, horseradish peroxidase, was studied in the presence of cellobiose oxidase (CbO) and cellobiose. It was found that the reversion rates for MnP compound II and lignin peroxidase compound II back to native enzymes increased significantly in the presence of CbO and cellobiose. However, the reduction of cytochrome c by CbO plus cellobiose was 40 times faster than the reduction of MnP compound II. Also, the lag phase before reversion to the native states decreased for all three peroxidases in the presence of CbO and cellobiose. Active CbO did not repress formation of compounds I or II of the peroxidases, and Mn2+/veratryl alcohol reduced compound II of the peroxidases much more rapidly than did active CbO. This indicates that, in the presence of Mn2+ or veratryl alcohol, MnP and lignin peroxidase can complete their catalytic cycles and function normally without interference from CbO. Without the presence of peroxidase substrates, active CbO reduced compound II of the above peroxidases.

scholarly journals Homologous Expression of Recombinant Lignin Peroxidase in Phanerochaete chrysosporium

1999 ◽  
Vol 65 (4) ◽  
pp. 1670-1674 ◽  
Author(s):  
Maarten D. Sollewijn Gelpke ◽  
Mary Mayfield-Gambill ◽  
Geoffrey P. Lin Cereghino ◽  
Michael H. Gold
Keyword(s):  
Phanerochaete Chrysosporium ◽  
Lignin Peroxidase ◽  
Active Form ◽  
Schizophyllum Commune ◽  
Transient State ◽  
Veratryl Alcohol ◽  
Kinetic Properties ◽  
Gene Encoding ◽  
Homologous Expression ◽  
State Kinetic

ABSTRACT The glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter was used to drive expression of lip2, the gene encoding lignin peroxidase (LiP) isozyme H8, in primary metabolic cultures of Phanerochaete chrysosporium. The expression vector, pUGL, also contained the Schizophyllum commune ura1gene as a selectable marker. pUGL was used to transform a P. chrysosporium Ura11 auxotroph to prototrophy. Ura+transformants were screened for peroxidase activity in liquid cultures containing high-carbon and high-nitrogen medium. Recombinant LiP (rLiP) was secreted in active form by the transformants after 4 days of growth, whereas endogenous lip genes were not expressed under these conditions. Approximately 2 mg of homogeneous rLiP/liter was obtained after purification. The molecular mass, pI, and optical absorption spectrum of rLiPH8 were essentially identical to those of the wild-type LiPh8 (wt LiPH8), indicating that heme insertion, folding, and secretion functioned normally in the transformant. Steady-state and transient-state kinetic properties for the oxidation of veratryl alcohol between wtLiPH8 and rLiPH8 were also identical.

Detection and Characterization of the Lignin Peroxidase Compound II−Veratryl Alcohol Cation Radical Complex†

Biochemistry ◽  
1997 ◽  
Vol 36 (46) ◽  
pp. 14181-14185 ◽  
Author(s):  
Aditya Khindaria ◽  
Guojun Nie ◽  
Steven D. Aust
Keyword(s):  
Lignin Peroxidase ◽  
Cation Radical ◽  
Veratryl Alcohol ◽  
Radical Complex ◽  
Compound Ii

Kinetic and Thermodynamic Characterization of Lignin Peroxidase Isolated from Agaricus bitorqus A66

2012 ◽  
Vol 10 (1) ◽  
Author(s):  
Ismat Bibi ◽  
Haq Nawaz Bhatti
Keyword(s):  
Lignin Peroxidase ◽  
Thermal Inactivation ◽  
Specific Activity ◽  
Gel Filtration ◽  
Veratryl Alcohol ◽  
Exchange Chromatography ◽  
Different Temperatures ◽  
Scale Experiment ◽  
Menten Kinetic

This study deals with purification and characterization of lignin peroxidase (LiP) isolated from Agaricus bitorqus A66 during decolorization of NOVASOL Direct Black dye. A laboratory scale experiment was conducted for maximum LiP production under optimal conditions. Purification & fractionation of LiP was performed on DEAE-Sepharose ion exchange chromatography followed by Sephadex G-50 gel filtration. The purified LiP has a specific activity of 519 U/mg with 6.73% activity recover. The optimum pH and temperature of purified LiP for the oxidation of veratryl alcohol were 6.8 and 45 °C, respectively. Michaelis-Menten kinetic constants (Vmax and Km) were determined using different concentrations of veratryl alcohol (1-35 mM). The Km and Vmax were 16.67 mM and 179.2 U/mL respectively, for veratryl alcohol oxidation as determined from the Lineweaver-Burk plot. Thermal inactivation studies were carried out at different temperatures to check the thermal stability of the enzyme. Enthalpy of activation decreased where Free energy of activation for thermal denaturation increased at higher temperatures. A possible explanation for the thermal inactivation of LiP at higher temperatures is also discussed.

Synthesis and application of Co(salen) complexes containing proximal imidazolium ionic liquid cores

2012 ◽  
Vol 90 (1) ◽  
pp. 60-70 ◽  
Author(s):  
Swapnil Sonar ◽  
Kenson Ambrose ◽  
Arthur D. Hendsbee ◽  
Jason D. Masuda ◽  
Robert D. Singer
Keyword(s):  
Ionic Liquid ◽  
Ionic Liquids ◽  
Model Compound ◽  
Veratryl Alcohol ◽  
Oxidation Catalysts ◽  
Imidazolium Ionic Liquid ◽  
Lignin Model Compound ◽  
Salen Complexes ◽  
Lignin Model ◽  
Salen Ligand

Ionic ligands derived from a salen ligand containing two proximal 1,3-disubstituted imidazolium ionic liquid cores form cobalt(III) complexes capable of selectively oxidizing veratryl alcohol, a lignin model compound, to veratraldehyde using air as the source of oxygen. These complexes are easy to prepare, inexpensive, water stable, and soluble in ionic liquids, making them viable candidates for use as oxidation catalysts.

scholarly journals Functional Characterization of Melanin Decolorizing Extracellular Peroxidase of Bjerkandera adusta

2021 ◽  
Vol 7 (9) ◽  
pp. 762
Author(s):  
Jina Baik ◽  
Anwesha Purkayastha ◽  
Kyung Hye Park ◽  
Taek Jin Kang
Keyword(s):  
Lignin Peroxidase ◽  
Functional Characterization ◽  
Veratryl Alcohol ◽  
Storage Conditions ◽  
Versatile Peroxidase ◽  
Bjerkandera Adusta ◽  
Cellular Mechanisms ◽  
Melanin Pigmentation ◽  
Crude Enzyme Preparation

Melanin pigmentation in the human skin results from complicated cellular mechanisms that remain to be entirely understood. Uneven melanin pigmentation has been counteracted by inhibiting synthesis or transfer of melanin in the skin. Recently, an enzymatic approach has been proposed, wherein the melanin in the skin is decolorized using lignin peroxidase. However, not many enzymes are available for decolorizing melanin; the most studied one is lignin peroxidase derived from a lignin degrading fungus, Phanerochaete chrysosporium. Our current study reveals that versatile peroxidase from Bjerkandera adusta can decolorize synthetic melanin. Melanin decolorization was found to be dependent on veratryl alcohol and hydrogen peroxide, but not on Mn2+. The degree of decolorization reached over 40% in 10 min at 37 °C and a pH of 4.5. Optimized storage conditions were slightly different from those for the reaction; crude enzyme preparation was the most stable at 25 °C at pH 5.5. Since the enzyme rapidly lost its activity at 50 °C, stabilizers were screened. As a result, glycerol, a major component in several cosmetic formulations, was found to be a promising excipient. Our results suggest that B. adusta versatile peroxidase can be considered for future cosmetic applications aimed at melanin decolorization.

Sign in / Sign up

Export Citation Format

Share Document