Screening of monokaryotic strains derived from Pleurotus sapidus for oxidoreductase enzymes production with biotechnological interest

FERNÁNDEZ-LAHORE M; LANDACHE M I; LOPRETTI M.; ZYGADLO J.; OMARINI A.

Murcia

Congreso; CONGRESO NACIONAL DE BIOTECNOLOGÍA (BIOTEC 2017); 2017

Sociedad Española de Biotecnología

A large number of fungal strains have been used to upgrade different agroindustry sise-streams through the production of enzymes and novel types of compounds by fermentation. Among them, basidiomycetes have shown the capability to produce oxidoreductase enzymes, laccase, and peroxidases, able to transform a broad range of organic compounds, such as polyphenols or terpenes present in the plant tissues. For example, Pleurotus sapidus (mono- and dikaryon strains) offer many technical advantages and have been used to produce a wide variety of enzymes or metabolites with industrial applications. Environmental tolerance is a complex and poorly understood phenotype including tolerance to products, substrates, by-products or bioprocess conditions. The genotype diversity of the (sexual) basidiospores (monokaryons, Mks) after meiosis will guarantee broad phenotype diversity, from which tolerant Mks (germinated spores) able to produce higher titers of oxidoreductase enzyme(s) can be selected. Spores obtained from the dikaryotic strains of P. sapidus Dk3174 were used to screen, on agar planes, Mks able to grow and degrade selective media supplemented with Remallo Brilliant Blue R (RBBR). 70 Mks strains able to degrade the RBBR, by the presence of oxidoreductase enzymes, were used in a second screening, on agar plates, to test the enzymes activities directly on fungal colonies using ABTS, guaiacol, cathecol, 2,6-dimethoxypehnol (Syringol), and veratryl alcohol. These substratos are oxidized by laccase, and by generic peroxidases, MnP, DyP, and VP in the presence of H2O2. As a result, it was possible to obtain strains with different pateras of enzymatic activity (Fig. 1), some of them with a predominance of the laccase or peroxidases, others with the presence of both but with different intensities and in other strains it was possible to detect VP. The relevante of this results lies in the use of a fast method to select Mks of P. sapidus with high viability and able to produceenzymes that can be used to transform polyphenols and terpenes, present in different Agroindustrial sise-streams (e.g. cereals, Citrus), into new bioactive compounds.