Kinetic and Structural Properties of a Robust Bacterial L-Amino Acid Oxidase

Catalysts

Simone Savino¹  Hugo L. van Beek¹  J. Daniël-Moráh Meijer²  Marco W. Fraaije²  Henriëtte J. Rozeboom² 

[1] GECCO Biotech, Nijenborgh 4, 9747 AG Groningen, The Netherlands;Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands;

关键词: flavin-dependent oxidase;    L-amino acids;    deracemisation;    biocatalysis;    crystal structure;   

DOI  :  10.3390/catal11111309

来源: DOAJ

【 摘 要 】

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 previously reported LAAO from the bacterium Pseudoalteromonas luteoviolacea. The substrate scope and kinetic properties of the enzyme were determined, and the thermostability was evaluated. Additionally, 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 expression of this thermostable LAAO provides a solid basis for the development of engineered LAAO variants tuned for biosensing and/or biocatalysis.

【 授权许可】

Unknown