Vector Engineering Anomalies: Impact on Fusion Protein Purification Performance

Richard S. Pasquinelli; Richard R. Koepsel; Naxin Wu; Mohammad M. Ataai

doi:10.1006/prep.1999.1152

Vector Engineering Anomalies: Impact on Fusion Protein Purification Performance

Protein Expression and Purification ◽

10.1006/prep.1999.1152 ◽

1999 ◽

Vol 17 (3) ◽

pp. 449-455 ◽

Cited By ~ 1

Author(s):

Richard S. Pasquinelli ◽

Richard R. Koepsel ◽

Naxin Wu ◽

Mohammad M. Ataai

Keyword(s):

Fusion Protein ◽

Protein Purification

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GST-fusion Protein Purification

SciVee ◽

10.4016/16485.01 ◽

2010 ◽

Author(s):

Abnova Corp

Keyword(s):

Fusion Protein ◽

Protein Purification ◽

Gst Fusion Protein

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

International Journal of Molecular Sciences ◽

10.3390/ijms21020416 ◽

2020 ◽

Vol 21 (2) ◽

pp. 416

Author(s):

Angel De La Cruz Pech-Canul ◽

Javier Carrillo-Campos ◽

María de Lourdes Ballinas-Casarrubias ◽

Rosa Lidia Solis-Oviedo ◽

Selena Karina Hernández-Rascón ◽

...

Keyword(s):

Fusion Protein ◽

Protein Purification ◽

Phanerochaete Chrysosporium ◽

Manganese Peroxidase ◽

Expression System ◽

White Rot Fungi ◽

Functional Expression ◽

White Rot ◽

E Coli ◽

One Step

Manganese peroxidases (MnP) from the white-rot fungi Phanerochaete chrysosporium catalyse the oxidation of Mn2+ to Mn3+, 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 P. chrysosporium using an E. coli-expression system. The proposed strategy involved the codon optimization and chemical synthesis of the MnP1 gene for optimised expression in the E. coli 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 Mn2+.

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