An engineered Phlebia radiata manganese peroxidase: expression, refolding, purification and preliminary characterization

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dc.contributor.authorF. Ufot, Usenobong
dc.contributor.authorAkpanabiatu, Monday I.
dc.date.accessioned2016-10-25T06:00:03Z
dc.date.available2016-10-25T06:00:03Z
dc.date.issued2012-10
dc.description.abstractManganese peroxidases (MnPs) are interesting enzymes in protein engineering, aimed at maximizing industrial bioprocesses such as lignin degradation and biofuel production. cDNA of the secreted short-type of MnP from Phlebia radiata (Pr-MnP3) has been successfully engineered and amplified by polymerase chain reaction (PCR). Five mutant genes (E40H, E44H, E40H/E44H, D186H and D186N) of recombinant Phlebia radiata MnP3 (rPr-MnP3) were generated. The wild-type and the mutant genes were expressed in Escherichia coli (W3110 strain) and the resultant body proteins were lysed, purified and refolded into active enzymes. 6% - 7% recovery of pure and fully active rPr-MnP3 for wild-type and mutants were obtained and the availability of rPr-MnP3 enzymes will greatly facilitate its structure-function relationships studies. rPr-MnP3 mass was characterised using SDS-PAGE and MALDI-TOF mass spectrometry. Molecular weight of both the wild-type and mutant rPr-MnP3 enzymes was approximately 36 kDa. This describes the spectral characterization of the wild-type and mutant rPr-MnP3 enzymes with are very close similarities; substantially high spin haem enzymes. Therefore we report the engineering, cloning, expression, refolding/activation of MnP3 genes and preliminary characterization of the wild-type and mutant Phlebia radiata MnP3 enzymes.en_US
dc.identifier.citationAmerican Journal of Molecular Biology, 2012, 2, 359-370en_US
dc.identifier.urihttp://dx.doi.org/10.4236/ajmb.2012.24037
dc.identifier.urihttp://hdl.handle.net/123456789/1023
dc.language.isoenen_US
dc.publisherScientific Research Publishingen_US
dc.subjectPhlebia Radiataen_US
dc.subjectPeroxidaseen_US
dc.subjectManganese Peroxidaseen_US
dc.subjectOver-Expressionen_US
dc.subjectRecombinanten_US
dc.titleAn engineered Phlebia radiata manganese peroxidase: expression, refolding, purification and preliminary characterizationen_US
dc.typeArticleen_US
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