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Engineering of pyranose 2-oxidase: Improvement for biofuel cell and food applications through semi-rational protein design by Oliver Spadiut; Ines Pisanelli; Thomas Maischberger; Clemens Peterbauer; Lo Gorton; Pimchai Chaiyen; Dietmar Haltrich is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is Engineering of pyranose 2-oxidase: Improvement for biofuel cell and food applications through semi-rational protein design about?
Pyranose 2-oxidase (P2Ox) has several proposed biotechnological applications such as a bio-component in biofuel cells or for carbohydrate transformations. To improve some of the catalytic properties of P2Ox from Trametes multicolor, we selected a semi-rational approach of enzyme engineering, saturation mutagenesis of active-site residues and subsequent screening of mutant libraries for improved activity. One of the active-site mutants with improved catalytic characteristics identified was V546C, which showed catalytic constants increased by up to 5.7-fold for both the sugar substrates (d-glucose and d-galactose) and alternative electron acceptors (1,4-benzoquinone, BQ and ferricenium ion, Fc + ], albeit at the expense of increased Michaelis constants. By combining V546C with other amino acid replacements, we obtained P2Ox variants that are of interest for biofuel cell applications due to their increased k cat for both BQ and Fc + , e.g., V546C/E542K showed 4.4-and 17-fold increased k cat for BQ compared to the wild-type enzyme when d-glucose and d-galactose, respectively, were the saturating substrates, while V546C/T169G showed approx. 40-and 50-fold higher k cat for BQ and Fc + ,
Who reads Engineering of pyranose 2-oxidase: Improvement for biofuel cell and food applications through semi-rational protein design?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Oliver Spadiut; Ines Pisanelli; Thomas Maischberger; Clemens Peterbauer; Lo Gorton; Pimchai Chaiyen; Dietmar Haltrich
- Publisher
- Elsevier Science; Elsevier ; Elsevier BV (ISSN 0168-1656)
- Published
- 2009
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)