Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity
| Autor: | Kyle J. Mamounis, Maria Luiza Caldas Nogueira, Daniela Priscila Marchi Salvador, Andres Andreo-Vidal, Antonio Sanchez-Amat, Victor L. Davidson |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Molecules, Vol 27, Iss 15, p 4726 (2022) |
| Druh dokumentu: | article |
| ISSN: | 27154726 1420-3049 |
| DOI: | 10.3390/molecules27154726 |
| Popis: | The Pseudoalteromonas luteoviolacea strain CPMOR-1 expresses a flavin adenine dinucleotide (FAD)-dependent L-amino acid oxidase (LAAO) with broad substrate specificity. Steady-state kinetic analysis of its reactivity towards the 20 proteinogenic amino acids showed some activity to all except proline. The relative specific activity for amino acid substrates was not correlated only with Km or kcat values, since the two parameters often varied independently of each other. Variation in Km was attributed to the differential binding affinity. Variation in kcat was attributed to differential positioning of the bound substrate relative to FAD that decreased the reaction rate. A structural model of this LAAO was compared with structures of other FAD-dependent LAAOs that have different substrate specificities: an LAAO from snake venom that prefers aromatic amino acid substrates and a fungal LAAO that is specific for lysine. While the amino acid sequences of these LAAOs are not very similar, their overall structures are comparable. The differential activity towards specific amino acids was correlated with specific residues in the active sites of these LAAOs. Residues in the active site that interact with the amino and carboxyl groups attached to the α-carbon of the substrate amino acid are conserved in all of the LAAOs. Residues that interact with the side chains of the amino acid substrates show variation. This provides insight into the structural determinants of the LAAOs that dictate their different substrate preferences. These results are of interest for harnessing these enzymes for possible applications in biotechnology, such as deracemization. |
| Databáze: | Directory of Open Access Journals |
| Externí odkaz: |
|