title: Recombinant expression of four oxidoreductases in Phanerochaete chrysosporium improves degradation of phenolic and non-phenolic substrates.
abstract: 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.
authors.affiliation: Centro de Investigación y de Estudios Avanzados del IPN, Unidad Irapuato, Km. 9.6 Libramiento Norte, Carretera Irapuato-León, 36500 Irapuato, Guanajuato, Mexico.
authors.forename: Nancy
authors.initials: N
authors.lastname: Coconi-Linares
authors.affiliation: Instituto Tecnológico de Mérida, Av. Tecnológico, 97118 Mérida, Yucatán, Mexico.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Ortiz-Vázquez
authors.affiliation: Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Boulevard Juriquilla 3001, 76230 Querétaro, Mexico.
authors.forename: Francisco
authors.initials: F
authors.lastname: Fernández
authors.affiliation: Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Boulevard Juriquilla 3001, 76230 Querétaro, Mexico; División de Ciencias de la Salud, Universidad del Valle de México, Villas del Mesón 1000, 76230 Querétaro, Mexico.
authors.forename: Achim M
authors.initials: AM
authors.lastname: Loske
authors.affiliation: Centro de Investigación y de Estudios Avanzados del IPN, Unidad Irapuato, Km. 9.6 Libramiento Norte, Carretera Irapuato-León, 36500 Irapuato, Guanajuato, Mexico. Electronic address: mgomez@ira.cinvestav.mx.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Gómez-Lim
mesh_terms: D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D042845:Laccase / Q000235:genetics / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods; D010544:Peroxidases / Q000235:genetics / Q000378:metabolism*; D020075:Phanerochaete / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D010636:Phenols / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D005656:Fungal Proteins; D010636:Phenols; D011994:Recombinant Proteins; D042845:Laccase; D010544:Peroxidases
keywords: Fungal oxidoreductases; Heterologous expression; Laccase; Phanerochaete chrysosporium; Phenolic compounds degradation; Shock waves