High-yield production of manganese peroxidase, lignin peroxidase, and versatile peroxidase in Phanerochaete chrysosporium
Por:
Coconi-Linares N., Magana-Ortiz, D, Guzmán-Ortiz D.A., Fernández F., Loske A.M., Gómez-Lim M.A.
Publicada:
1 nov 2014
Resumen:
The white-rot fungus Phanerochaete chrysosporium secretes extracellular
oxidative enzymes during secondary metabolism, but lacks versatile
peroxidase, an enzyme important in ligninolysis and diverse
biotechnology processes. In this study, we report the genetic
modification of a P. chrysosporium strain capable of co-expressing two
endogenous genes constitutively, manganese peroxidase (mnp1) and lignin
peroxidase (lipH8), and the codon-optimized vpl2 gene from Pleurotus
eryngii. For this purpose, we employed a highly efficient transformation
method based on the use of shock waves developed by our group. The
expression of recombinant genes was verified by PCR, Southern blot,
quantitative real-time PCR (qRT-PCR), and assays of enzymatic activity.
The production yield of ligninolytic enzymes was up to four times higher
in comparison to previously published reports. These results may
represent significant progress toward the stable production of
ligninolytic enzymes and the development of an effective fungal strain
with promising biotechnological applications.
Filiaciones:
Coconi-Linares N.:
Centro de Investigación y de Estudios Avanzados del IPN, Unidad Irapuato, Km. 9.6 Carretera Irapuato-León, Irapuato, Gto 36821, Mexico
Guzmán-Ortiz D.A.:
Centro de Investigación y de Estudios Avanzados del IPN, Unidad Irapuato, Km. 9.6 Carretera Irapuato-León, Irapuato, Gto 36821, Mexico
Fernández F.:
Univ Nacl Autonoma Mexico, Ctr Fis Aplicada & Tecnol Avanzada, Queretaro 76000, Qro, Mexico
Loske A.M.:
Univ Nacl Autonoma Mexico, Ctr Fis Aplicada & Tecnol Avanzada, Queretaro 76000, Qro, Mexico
Gómez-Lim M.A.:
Centro de Investigación y de Estudios Avanzados del IPN, Unidad Irapuato, Km. 9.6 Carretera Irapuato-León, Irapuato, Gto 36821, Mexico
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