Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic Acids
The oxidation of alcohols to the corresponding carbonyl or carboxyl compounds represents a convenient strategy for the selective introduction of electrophilic carbon centres into carbohydrate-based starting materials. The O2-dependent oxidation of prim-alcohols by flavin-containing alcohol oxidases...
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| Format: | Article |
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MDPI AG
2017-12-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/22/12/2205 |
| _version_ | 1817976919198531584 |
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| author | Mathias Pickl Christoph K. Winkler Silvia M. Glueck Marco W. Fraaije Kurt Faber |
| author_facet | Mathias Pickl Christoph K. Winkler Silvia M. Glueck Marco W. Fraaije Kurt Faber |
| author_sort | Mathias Pickl |
| collection | DOAJ |
| description | The oxidation of alcohols to the corresponding carbonyl or carboxyl compounds represents a convenient strategy for the selective introduction of electrophilic carbon centres into carbohydrate-based starting materials. The O2-dependent oxidation of prim-alcohols by flavin-containing alcohol oxidases often yields mixtures of aldehyde and carboxylic acid, which is due to “over-oxidation” of the aldehyde hydrate intermediate. In order to directly convert alcohols into carboxylic acids, rational engineering of 5-(hydroxymethyl)furfural oxidase was performed. In an attempt to improve the binding of the aldehyde hydrate in the active site to boost aldehyde-oxidase activity, two active-site residues were exchanged for hydrogen-bond-donating and -accepting amino acids. Enhanced over-oxidation was demonstrated and Michaelis–Menten kinetics were performed to corroborate these findings. |
| first_indexed | 2024-04-13T22:09:13Z |
| format | Article |
| id | doaj.art-a8ce64438a014982852fc793f5b01550 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-04-13T22:09:13Z |
| publishDate | 2017-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-a8ce64438a014982852fc793f5b015502022-12-22T02:27:48ZengMDPI AGMolecules1420-30492017-12-012212220510.3390/molecules22122205molecules22122205Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic AcidsMathias Pickl0Christoph K. Winkler1Silvia M. Glueck2Marco W. Fraaije3Kurt Faber4Department of Chemistry, University of Graz, Heinrichstrasse 28, A-8010 Graz, AustriaAustrian Centre of Industrial Biotechnology, ACIB GmbH c/o Department of Chemistry, University of Graz, Heinrichstrasse 28, A-8010 Graz, AustriaAustrian Centre of Industrial Biotechnology, ACIB GmbH c/o Department of Chemistry, University of Graz, Heinrichstrasse 28, A-8010 Graz, AustriaMolecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The NetherlandsDepartment of Chemistry, University of Graz, Heinrichstrasse 28, A-8010 Graz, AustriaThe oxidation of alcohols to the corresponding carbonyl or carboxyl compounds represents a convenient strategy for the selective introduction of electrophilic carbon centres into carbohydrate-based starting materials. The O2-dependent oxidation of prim-alcohols by flavin-containing alcohol oxidases often yields mixtures of aldehyde and carboxylic acid, which is due to “over-oxidation” of the aldehyde hydrate intermediate. In order to directly convert alcohols into carboxylic acids, rational engineering of 5-(hydroxymethyl)furfural oxidase was performed. In an attempt to improve the binding of the aldehyde hydrate in the active site to boost aldehyde-oxidase activity, two active-site residues were exchanged for hydrogen-bond-donating and -accepting amino acids. Enhanced over-oxidation was demonstrated and Michaelis–Menten kinetics were performed to corroborate these findings.https://www.mdpi.com/1420-3049/22/12/2205biocatalysisalcohol oxidationaldehyde oxidationflavoprotein oxidaseprotein design |
| spellingShingle | Mathias Pickl Christoph K. Winkler Silvia M. Glueck Marco W. Fraaije Kurt Faber Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic Acids Molecules biocatalysis alcohol oxidation aldehyde oxidation flavoprotein oxidase protein design |
| title | Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic Acids |
| title_full | Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic Acids |
| title_fullStr | Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic Acids |
| title_full_unstemmed | Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic Acids |
| title_short | Rational Engineering of a Flavoprotein Oxidase for Improved Direct Oxidation of Alcohols to Carboxylic Acids |
| title_sort | rational engineering of a flavoprotein oxidase for improved direct oxidation of alcohols to carboxylic acids |
| topic | biocatalysis alcohol oxidation aldehyde oxidation flavoprotein oxidase protein design |
| url | https://www.mdpi.com/1420-3049/22/12/2205 |
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