Copper is an essential trace element in living systems, present in the parts per million concentration range. It is a key cofactor in a diverse array of biological oxidation-reduction reactions. These involve either outer-sphere electron transfer, as in the blue copper proteins and the Cu{sub A} site of cytochrome oxidase and nitrous oxide redutase, or inner-sphere electron transfer in the binding, activation, and reduction of dioxygen, superoxide, nitrite, and nitrous oxide. Copper sites have historically been divided into three classes based on their spectroscopic features, which reflect the geometric and electronic structure of the active site: type 1 (T1) or blue copper, type 2 (T2) or normal copper, and type 3 (T3) or coupled binuclear copper centers. 428 refs.

Multicopper Oxidases and Oxygenases

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The structure and function of fungal laccases

Cellulases and related enzymes in biotechnology

White-rot fungi produce extracellular lignin-modifying enzymes, the best characterized of which are laccase (EC 1.10.3.2), lignin peroxidases (EC 1.11.1.7) and manganese peroxidases (EC 1.11.1.7). Lignin biodegradation studies have been carried out mostly using the white-rot fungus Phanerochaete chrysosporium which produces multiple isoenzymes of lignin peroxidase and manganese peroxidase but does not produce laccase. Many other white-rot fungi produce laccase in addition to lignin and manganese peroxidases and in varying combinations. Based on the enzyme production patterns of an array of white-rot fungi, three categories of fungi are suggested: (i) lignin-manganese peroxidase group (e.g.P. chrysosporium and Phlebia radiata), (ii) manganese peroxidase-laccase group (e.g. Dichomitus squalens and Rigidoporus lignosus), and (iii) lignin peroxidase-laccase group (e.g. Phlebia ochraceofulva and Junghuhnia separabilima). The most efficient lignin degraders, estimated by 14CO2 evolution from 14C-[Ring]-labelled synthetic lignin (DHP), belong to the first group, whereas many of the most selective lignin-degrading fungi belong to the second, although only moderate to good [14C]DHP mineralization is obtained using fungi from this group. The lignin peroxidase-laccase fungi only poorly degrade [14C]DHP.

Lignin-modifying enzymes from selected white-rot fungi: production and role from in lignin degradation

Using enzymes as decontaminating agents has received great attention. One of the most promising groups of enzymes, laccases, are used to decontaminate phenol-polluted systems and for bio technological applications. Higher plants and fungi, mostly wood-rotting fungi, are the main producers of laccases, but bacterial laccases also have been found. Belonging to the class of phenoloxidases, laccases catalyze the polymerization of several phenolic substances to polymeric products. In addition, they have transformed lignin and lignin-related compounds, showing a very broad substrate specificity. Specific compounds acting as protein-synthesis inducers historically have been used to improve the production of the enzyme. Recent success in fungal molecular and cellular engineering technology has contributed to significantly increase the industrial production of recombinant laccase. Kinetic (Michaelis-Menten parameters, optimum pH, kcat) and stability properties of laccases may vary according to the source of the en...

https://www.tandfonline.com/doi/pdf/10.1080/10889869991219163

Laccases: A Useful Group of Oxidoreductive Enzymes

Summary: We have isolated and characterized a gene coding for the laccase of the lignin-degrading fungus Phlebia radiata. The gene has nine introns and recognizable fungal promoter elements. Sequences homologous to the consensus eukaryotic heat-shock regulatory element can be found in the promoter. RNA hybridization results indicate that this gene is regulated at the transcriptional level. The derived laccase amino acid sequence shows homology to plant ascorbate oxidases, suggesting that the basic structure of the laccase is similar to the three-fold repeated β-barrel of the ascorbate oxidases. Potential copper ligands and a residue carrying the prosthetic group pyrroloquinoline quinone (PQQ) in the laccase protein can be identified by homology. The intron/exon structure of the laccase gene suggests that this protein could have evolved by exon shuffling.

Isolation and structural analysis of the laccase gene from the lignin degrading fungus Phlebia radiata

We have expressed a gene encoding the ligninolytic laccase enzyme of the white–rot fungus Phlebia radiata in the soft rot fungus Trichoderma reesei under the promoter of the major cellulase gene (cbh1). Constructions from the laccase cDNA or the chromosomal gene were equivalent in expression, yielding 20 mg/l of secreted active laccase in small–scale fermentations. The recombinant enzyme has a similar molecular weight, antigenic properties and specific activity as the Phlebia laccase and is analogous to the Phlebia enzyme in removing monomeric lignin–related compounds from residual lignin of the pulping process, thus demonstrating its potential for industrial applications.

Marja-Leena Niku-Paavola

Papers

Isolation and structural analysis of the laccase gene from the lignin degrading fungus Phlebia radiata

Heterologous Production of a Ligninolytic Enzyme: Expression of the Phlebia Radiata Laccase Gene in Trichoderma Reesei

A lignin peroxidase-encoding cDNA from the white-rot fungus Phlebia radiata: characterization and expression in Trichoderma reesei.