Discovery of Two Novel Oxidases Using a High-Throughput Activity Screen
Journal article, 2022

Discovery of novel enzymes is a challenging task, yet a crucial one, due to their increasing relevance as chemical catalysts and biotechnological tools. In our work we present a high-throughput screening approach to discovering novel activities. A screen of 96 putative oxidases with 23 substrates led to the discovery of two new enzymes. The first enzyme, N-acetyl-D-hexosamine oxidase (EC 1.1.3.29) from Ralstonia solanacearum, is a vanillyl alcohol oxidase-like flavoprotein displaying the highest activity with N-acetylglucosamine and N-acetylgalactosamine. Before our discovery of the enzyme, its activity was an orphan one - experimentally characterized but lacking the link to amino acid sequence. The second enzyme, from an uncultured marine euryarchaeota, is a long-chain alcohol oxidase (LCAO, EC 1.1.3.20) active with a range of fatty alcohols, with 1-dodecanol being the preferred substrate. The enzyme displays no sequence similarity to previously characterised LCAOs, and thus is a completely novel representative of a protein with such activity.

flavoproteins

high-throughput screening

oxidoreductases

enzyme discovery

orphan enzymes

Author

Elzbieta Rembeza

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Alessandro Boverio

University of Groningen

Marco W. Fraaije

University of Groningen

Martin Engqvist

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

ChemBioChem

1439-4227 (ISSN) 1439-7633 (eISSN)

Vol. 23 2 e202100510

Subject Categories

Biochemistry and Molecular Biology

Structural Biology

Biocatalysis and Enzyme Technology

DOI

10.1002/cbic.202100510

PubMed

34709726

More information

Latest update

4/5/2022 6