Recombinant expression of four oxidoreductases in Phanerochaete chrysosporium improves degradation of phenolic and non-phenolic substrates

• Published in: Journal of biotechnology Vol. 209; pp. 76 - 84
• Main Authors: Coconi-Linares, Nancy, Ortiz-Vázquez, Elizabeth, Fernández, Francisco, Loske, Achim M., Gómez-Lim, Miguel A.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.09.2015
• Subjects: Basidiomycetes; Cloning, Molecular; Enzymes; Fungal oxidoreductases; Fungal Proteins - genetics; Fungal Proteins - metabolism; Fungi; Genes; Heterologous expression; Laccase; Laccase - genetics; Laccase - metabolism; Metabolic Engineering - methods; Peroxidase; Peroxidases - genetics; Peroxidases - metabolism; Phanerochaete - enzymology; Phanerochaete - genetics; Phanerochaete - metabolism; Phanerochaete chrysosporium; Phenolic compounds degradation; Phenols - metabolism; Pleurotus eryngii; Recombinant; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Shock waves; Strain; Trametes versicolor; Transformation, Genetic; Phenolic compounds degradation; Shock waves; Phanerochaete chrysosporium; Fungal oxidoreductases; Laccase; Heterologous expression
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2015.06.401

•Four oxidoreductases were simultaneously co-expressed in P. chrysosporium.•The recombinant strains were able to produce VPL2 and LACIIIb in functional form.•Recombinant strains showed a much greater degradative capacity than the wild type.•A synergy between native and recombinant enzymes in substrate degradation was found. Phanerochaete chrysosporium belongs to a group of lignin-degrading fungi that secretes various oxidoreductive enzymes, including lignin peroxidase (LiP) and manganese peroxidase (MnP). Previously, we demonstrated that the heterologous expression of a versatile peroxidase (VP) in P. chrysosporium recombinant strains is possible. However, the production of laccases (Lac) in this fungus has not been completely demonstrated and remains controversial. In order to investigate if the co-expression of Lac and VP in P. chrysosporium would improve the degradation of phenolic and non-phenolic substrates, we tested the constitutive co-expression of the lacIIIb gene from Trametes versicolor and the vpl2 gene from Pleurotus eryngii, and also the endogenous genes mnp1 and lipH8 by shock wave mediated transformation. The co-overexpression of peroxidases and laccases was improved up to five-fold as compared with wild type species. Transformant strains showed a broad spectrum in phenolic/non-phenolic biotransformation and a high percentage in synthetic dye decolorization in comparison with the parental strain. Our results show that the four enzymes can be constitutively expressed in a single transformant of P. chrysosporium in minimal medium. These data offer new possibilities for an easy and efficient co-expression of laccases and peroxidases in suitable basidiomycete species.