Kinetic and structural properties of a robust bacterial l‐ amino acid oxidase

Simone Savino, J. Daniël‐Moráh Meijer, Henriëtte J. Rozeboom, Hugo L. van Beek, Marco W. Fraaije*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

L‐Amino acid oxidase (LAAO) is a flavin adenine dinucleotide (FAD)‐dependent enzyme active on most proteinogenic L‐amino acids, catalysing their conversion to α‐keto acids by oxidative deamination of the substrate. For this oxidation reaction, molecular oxygen is used as the electron acceptor, generating hydrogen peroxide. LAAO can be used to detect L‐amino acids, for the production of hydrogen peroxide as an oxidative agent or antimicrobial agent, and for the production of enantiopure amino acids from racemates. In this work, we characterised a previous-ly reported LAAO from the bacterium Pseudoalteromonas luteoviolacea. The substrate scope and kinetic properties of the enzyme were determined, and the thermostability was evaluated. Addi-tionally, we elucidated the crystal structure of this bacterial LAAO, enabling us to test the role of active site residues concerning their function in catalysis. The obtained insights and ease of ex-pression of this thermostable LAAO provides a solid basis for the development of engineered LAAO variants tuned for biosensing and/or biocatalysis.

Original languageEnglish
Article number1309
Number of pages9
JournalCatalysts
Volume11
Issue number11
DOIs
Publication statusPublished - Nov-2021

Keywords

  • Biocatalysis
  • Crystal structure
  • Deracemisation
  • Flavin‐dependent oxidase
  • L‐amino acids

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