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Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity
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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9331233/ https://www.ncbi.nlm.nih.gov/pubmed/35897902 http://dx.doi.org/10.3390/molecules27154726 |
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author | Mamounis, Kyle J. Caldas Nogueira, Maria Luiza Marchi Salvador, Daniela Priscila Andreo-Vidal, Andres Sanchez-Amat, Antonio Davidson, Victor L. |
author_facet | Mamounis, Kyle J. Caldas Nogueira, Maria Luiza Marchi Salvador, Daniela Priscila Andreo-Vidal, Andres Sanchez-Amat, Antonio Davidson, Victor L. |
author_sort | Mamounis, Kyle J. |
collection | PubMed |
description | 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 K(m) or k(cat) values, since the two parameters often varied independently of each other. Variation in K(m) was attributed to the differential binding affinity. Variation in k(cat) 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. |
format | Online Article Text |
id | pubmed-9331233 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-93312332022-07-29 Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity Mamounis, Kyle J. Caldas Nogueira, Maria Luiza Marchi Salvador, Daniela Priscila Andreo-Vidal, Andres Sanchez-Amat, Antonio Davidson, Victor L. Molecules Article 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 K(m) or k(cat) values, since the two parameters often varied independently of each other. Variation in K(m) was attributed to the differential binding affinity. Variation in k(cat) 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. MDPI 2022-07-24 /pmc/articles/PMC9331233/ /pubmed/35897902 http://dx.doi.org/10.3390/molecules27154726 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Mamounis, Kyle J. Caldas Nogueira, Maria Luiza Marchi Salvador, Daniela Priscila Andreo-Vidal, Andres Sanchez-Amat, Antonio Davidson, Victor L. Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity |
title | Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity |
title_full | Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity |
title_fullStr | Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity |
title_full_unstemmed | Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity |
title_short | Structural Determinants of the Specific Activities of an L-Amino Acid Oxidase from Pseudoalteromonas luteoviolacea CPMOR-1 with Broad Substrate Specificity |
title_sort | structural determinants of the specific activities of an l-amino acid oxidase from pseudoalteromonas luteoviolacea cpmor-1 with broad substrate specificity |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9331233/ https://www.ncbi.nlm.nih.gov/pubmed/35897902 http://dx.doi.org/10.3390/molecules27154726 |
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