dc.contributor.author | Lundell, Taina | |
dc.contributor.author | Bentley, Elodie | |
dc.contributor.author | Hilden, Kristiina | |
dc.contributor.author | Rytioja, Johanna | |
dc.contributor.author | Kuuskeri, Jaana | |
dc.contributor.author | Ufot, Usenobong F. | |
dc.contributor.author | Nousiainen, Paula | |
dc.contributor.author | Hofrichter, Martin | |
dc.contributor.author | Wahlsten, Matti | |
dc.contributor.author | Doyle, Wendy | |
dc.contributor.author | Smith, Andrew T. | |
dc.date.accessioned | 2017-05-22T02:50:58Z | |
dc.date.available | 2017-05-22T02:50:58Z | |
dc.date.issued | 2017 | |
dc.identifier.issn | 2211-551X | |
dc.identifier.doi | 10.2174/2211550105666160520120101 | |
dc.identifier.uri | http://hdl.handle.net/10072/337474 | |
dc.description.abstract | Background: Manganese peroxidases (MnP) and lignin peroxidases (LiP) are haem-including fungal secreted class-II peroxidases, which are interesting oxidoreductases in protein engineering aimed at designing of biocatalysts for lignin and lignocellulose conversion, dye compound degradation, activation of aromatic compounds, and biofuel production.
Objective. Recombinant short-type MnP (Pr-MnP3) of the white rot fungus Phlebia radiata, and its manganese- binding site (E40, E44, D186) directed variants were produced and characterized. To allow catalytic applications, enzymatic bleaching of Reactive Blue 5 and conversion of lignin-like compounds by engineered class- II peroxidases were explored.
Method: Pr-MnP3 and its variants were expressed in Escherichia coli. The resultant body proteins were lysed, purified and refolded into haem-including enzymes in 6-7% protein recovery, and examined spectroscopically and kinetically.
Results: Successful production of active enzymes was attained, with spectral characteristics of high-spin class-II peroxidases. Recombinant Pr-MnP3 demonstrated high affinity to Mn2+, which was noticeably affected by single (D186H/N) and double (E40H+E44H) mutations. Without addition of Mn2+, Pr- MnP3 was able to oxidize ABTS and decolorize Reactive Blue 5. Pc-LiPH8, its Trp-radical site variants, and engineered CiP-LiP demonstrated conversion of veratryl alcohol and dimeric non-phenolic lignin-model compounds (arylglycerol-β-aryl ethers) with production of veratraldehyde, which is evidence for cation radical formation with subsequent Cα-Cβ cleavage. Pc-LiPH8 and CiP variants were able to effectively oxidize and convert the phenolic dimer (guaiacylglycerol-β-guaiacyl ether).
Conclusion: Our results demonstrate suitability of engineered MnP and LiP peroxidases for dyedecolorizing, and efficiency of LiP and its variants for activation and degradation of phenolic and nonphenolic lignin-like aryl ether-linked compounds. | |
dc.description.peerreviewed | Yes | |
dc.language | English | |
dc.language.iso | eng | |
dc.publisher | Bentham Science | |
dc.relation.ispartofpagefrom | 116 | |
dc.relation.ispartofpageto | 127 | |
dc.relation.ispartofissue | 2 | |
dc.relation.ispartofjournal | Current Biotechnology | |
dc.relation.ispartofvolume | 6 | |
dc.subject.fieldofresearch | Medicinal and Biomolecular Chemistry not elsewhere classified | |
dc.subject.fieldofresearch | Biological Sciences | |
dc.subject.fieldofresearch | Technology | |
dc.subject.fieldofresearchcode | 030499 | |
dc.subject.fieldofresearchcode | 06 | |
dc.subject.fieldofresearchcode | 10 | |
dc.title | Engineering Towards Catalytic Use of Fungal Class-II Peroxidases for Dye-Decolorizing and Conversion of Lignin Model Compounds | |
dc.type | Journal article | |
dc.type.description | C1 - Articles | |
dc.type.code | C - Journal Articles | |
gro.hasfulltext | No Full Text | |
gro.griffith.author | Smith, Andrew T. | |