title: Heterologous expression of manganese peroxidase in Aspergillus niger and its effect on phenanthrene removal from soil.
abstract: A strain of Aspergillus niger, previously isolated from sugarcane bagasse because of its capacity to degrade phenanthrene in soil by solid culture, was used to express a manganese peroxidase gene (mnp1) from Phanerochaete chrysosporium, aiming at increasing its polycyclic aromatic hydrocarbons degradation capacity. Transformants were selected based on their resistance to hygromycin B and the discoloration induced on Poly R-478 dye by the peroxidase activity. The recombinant A. niger SBC2-T3 strain developed MnP activity and was able to remove 95% of the initial phenanthrene (400 ppm) from a microcosm soil system after 17 days, whereas the wild strain removed 72% under the same conditions. Transformation success was confirmed by PCR amplification using gene-specific primers, and a single fragment (1,348 bp long, as expected) of the recombinant mnp1 was amplified in the DNA from transformants, which was absent from the parental strain.
authors.affiliation: Centro de Investigación en Biotecnologóa Aplicada del IPN, Carretera Federal Santa Inés, Tepetitla de Lardizabal, Mexico.
authors.forename: Diana V
authors.initials: DV
authors.lastname: Cortés-Espinosa
authors.forename: Ángel E
authors.initials: ÁE
authors.lastname: Absalón
authors.forename: Noé
authors.initials: N
authors.lastname: Sanchez
authors.forename: Octavio
authors.initials: O
authors.lastname: Loera
authors.forename: Refugio
authors.initials: R
authors.lastname: Rodríguez-Vázquez
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Fernández
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D001711:Biotransformation; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D010544:Peroxidases / Q000235:genetics* / Q000378:metabolism*; D020075:Phanerochaete / Q000201:enzymology / Q000235:genetics; D010616:Phenanthrenes / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012641:Selection, Genetic; D012989:Soil Pollutants / Q000378:metabolism*; D013997:Time Factors; D014170:Transformation, Genetic
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D010616:Phenanthrenes; D011994:Recombinant Proteins; D012989:Soil Pollutants; C031181:phenanthrene; D010544:Peroxidases; C051129:manganese peroxidase