Functional Expression and One-Step Protein Purification of Manganese Peroxidase 1 (rMnP1) from Phanerochaete chrysosporium Using the E. coli -Expression System

Manganese peroxidases (MnP) from the white-rot fungi catalyse the oxidation of Mn to Mn , a strong oxidizer able to oxidize a wide variety of organic compounds. Different approaches have been used to unravel the enzymatic properties and potential applications of MnP. However, these efforts have been...

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Bibliographic Details
Published in:International journal of molecular sciences 2020-01, Vol.21 (2), p.416
Main Authors: Pech-Canul, Angel De La Cruz, Carrillo-Campos, Javier, Ballinas-Casarrubias, María de Lourdes, Solis-Oviedo, Rosa Lidia, Hernández-Rascón, Selena Karina, Hernández-Ochoa, León Raúl, Gutiérrez-Méndez, Néstor, García-Triana, Antonio
Format: Article
Language:English
Subjects:
Affinity chromatography
Chemical bonds
Communication
E coli
Enzymes
Fungi
Fusion protein
Gene expression
Guaiacol
Inclusion bodies
Lignin
Manganese
Organic compounds
Oxidation
Oxidizing agents
Peroxidase
Phanerochaete chrysosporium
Protein purification
Proteins
White rot
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Summary:Manganese peroxidases (MnP) from the white-rot fungi catalyse the oxidation of Mn to Mn , a strong oxidizer able to oxidize a wide variety of organic compounds. Different approaches have been used to unravel the enzymatic properties and potential applications of MnP. However, these efforts have been hampered by the limited production of native MnP by fungi. Heterologous expression of MnP has been achieved in both eukaryotic and prokaryotic expression systems, although with limited production and many disadvantages in the process. Here we described a novel molecular approach for the expression and purification of manganese peroxidase isoform 1 (MnP1) from using an -expression system. The proposed strategy involved the codon optimization and chemical synthesis of the MnP1 gene for optimised expression in the T7 shuffle host. Recombinant MnP1 (rMnP1) was expressed as a fusion protein, which was recovered from solubilised inclusion bodies. rMnP1 was purified from the fusion protein using intein-based protein purification techniques and a one-step affinity chromatography. The designated strategy allowed production of an active enzyme able to oxidize guaiacol or Mn .
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms21020416