Extension of Polyphenolics by CWPO-C Peroxidase Mutant Containing Radical-Robust Surface Active Site

Expressed as insoluble forms in Escherichia coli, native cationic cell wall peroxidase (CWPO-C) from the poplar tree and mutant variants were successfully reactivated via refolding experiments and used to elucidate the previously presumed existence of an electron transfer (ET) pathway in the CWPO-C...

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Bibliographic Details
Published in:	Applied biochemistry and biotechnology Vol. 172; no. 2; pp. 792 - 805
Main Authors:	Pham, L. T. Mai, Kim, S. Jin, Ahn, U. Suk, Choi, J. Weon, Song, B. Keun, Kim, Y. Hwan
Format:	Journal Article
Language:	English
Published:	Boston Springer-Verlag 2014 Springer US Springer Springer Nature B.V
Subjects:	antioxidant activity Antioxidants - metabolism Biocatalysis Biochemistry Biological and medical sciences Biotechnology Catalytic Domain catechin Catechin - chemistry Catechin - metabolism Cations Cell Wall - enzymology cell walls Chemistry Chemistry and Materials Science Chromatography, Gel Dimerization E coli electron transfer Electron transfer reactions Enzyme Stability Enzymes Escherichia coli Free Radicals - metabolism Fundamental and applied biological sciences. Psychology Green chemistry Hydrogen peroxide Kinetics Molecular Docking Simulation molecular weight Mutants Mutation - genetics Oxidation Oxidation-Reduction peroxidase Peroxidase - metabolism Phenols - chemistry Phenols - metabolism Polymerization Polymers Polyphenols Polyphenols - metabolism Populus - enzymology Spectrophotometry Structural Homology, Protein Thermodynamics tryptophan tyrosine xanthine oxidase Cell wall peroxidase poplar peroxidase Radical-robust Electron transfer mechanism Poly(catechin) Catechin Enzyme Active site Electron transfer Mechanism Cell wall S-peroxidase Peroxidases Peroxidase Oxidoreductases Mutation
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Summary:	Expressed as insoluble forms in Escherichia coli, native cationic cell wall peroxidase (CWPO-C) from the poplar tree and mutant variants were successfully reactivated via refolding experiments and used to elucidate the previously presumed existence of an electron transfer (ET) pathway in the CWPO-C structure. Their catalytic properties were fully characterized through various analyses including steady-state kinetic, direct oxidation of lignin macromolecules and their respective stabilities during the polymerization reactions. The analysis results proved that the 74th residue on the CWPO-C surface plays an important role in catalyzing the macromolecules via supposed ET mechanism. By comparing the residual activities of wild-type CWPO-C and mutant 74W CWPO-C after 3 min, mutation of tyrosine 74 residue to tryptophan increased the radical resistance of peroxidase up to ten times dramatically while maintaining its capability to oxidize lignin macromolecules. Furthermore, extension of poly(catechin) as well as lignin macromolecules with CWPO-C Y74W mutant clearly showed that this radical-resistant peroxidase mutant can increase the molecular weight of various kinds of polyphenolics by using surface-located active site. The anti-oxidation activity of the synthesized poly(catechin) was confirmed by xanthine oxidase assay. The elucidation of a uniquely catalytic mechanism in CWPO-C may improve the applicability of the peroxidase/H₂O₂ catalyst to green polymer chemistry.
Bibliography:	http://dx.doi.org/10.1007/s12010-013-0534-2 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0273-2289 1559-0291
DOI:	10.1007/s12010-013-0534-2