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Design of Artificial Alcohol Oxidases: Alcohol Dehydrogenase–NADPH Oxidase Fusions for Continuous Oxidations
- Source :
- Chembiochem, ChemBioChem, ChemBioChem, 20(1):cbic.201800421, 51-56. WILEY-V C H VERLAG GMBH
- Publication Year :
- 2018
- Publisher :
- John Wiley and Sons Inc., 2018.
-
Abstract
- To expand the arsenal of industrially applicable oxidative enzymes, fusions of alcohol dehydrogenases with an NADPH‐oxidase were designed. Three different alcohol dehydrogenases (LbADH, TbADH, ADHA) were expressed with a thermostable NADPH‐oxidase fusion partner (PAMO C65D) and purified. The resulting bifunctional biocatalysts retained the catalytic properties of the individual enzymes, and acted essentially like alcohol oxidases: transforming alcohols to ketones by using dioxygen as mild oxidant, while merely requiring a catalytic amount of NADP+. In small‐scale reactions, the purified fusion enzymes show good performances, with 69–99 % conversion, 99 % ee with a racemic substrate, and high cofactor and enzyme total turnover numbers. As the fusion enzymes essentially act as oxidases, we found that commonly used high‐throughput oxidase‐activity screening methods can be used. Therefore, if needed, the fusion enzymes could be easily engineered to tune their properties.<br />Going self‐sufficient: Alcohol dehydrogenases (ADHs) can catalyze selective alcohol oxidations and can be applied to kinetically resolve racemic alcohols, or to produce carbonyl building blocks. By fusing ADHs with an NADPH oxidase, self‐sufficient biocatalysts can be produced that act like alcohol oxidases. These enzyme fusions can be used for conversions and for activity screening to aid enzyme engineering.
- Subjects :
- biocatalysis
Recombinant Fusion Proteins
Levilactobacillus brevis
Alcohol
Thermoanaerobacter
010402 general chemistry
Protein Engineering
01 natural sciences
Biochemistry
Cofactor
Armoracia
Micrococcus
chemistry.chemical_compound
Very Important Paper
Oxidative enzyme
Escherichia coli
Animals
Bifunctional
Molecular Biology
Benzyl Alcohols
Alcohol dehydrogenase
chemistry.chemical_classification
alcohol dehydrogenases
biology
010405 organic chemistry
Communication
Organic Chemistry
Alcohol Dehydrogenase
NADPH Oxidases
Protein engineering
fusion enzymes
Cyclohexanols
Multifunctional Enzymes
Communications
0104 chemical sciences
enzyme engineering
Alcohol Oxidoreductases
Enzyme
chemistry
Biocatalysis
oxidases
biology.protein
Molecular Medicine
Cattle
Oxidation-Reduction
Subjects
Details
- Language :
- English
- ISSN :
- 14397633 and 14394227
- Volume :
- 20
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Chembiochem
- Accession number :
- edsair.doi.dedup.....869826bd6e607069b855b6544c7d3b75