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Purification, Characterization and Inhibition of Alanine Racemase from a Pathogenic Strain of Streptococcus iniae
Streptococcus iniae is a pathogenic and zoonotic bacteria that impacted high mortality to many fish species as well as capable of causing serious disease to humans. Alanine racemase (Alr, EC 5.1.1.1) is a pyridoxal-5’-phosphate (PLP)-containing homodimeric enzyme that catalyzes the racemization of L...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Exeley Inc.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7256847/ https://www.ncbi.nlm.nih.gov/pubmed/31880879 http://dx.doi.org/10.33073/pjm-2019-036 |
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author | MUHAMMAD, MURTALA LI, YANGYANG GONG, SIYU SHI, YANMIN JU, JIANSONG ZHAO, BAOHUA LIU, DONG |
author_facet | MUHAMMAD, MURTALA LI, YANGYANG GONG, SIYU SHI, YANMIN JU, JIANSONG ZHAO, BAOHUA LIU, DONG |
author_sort | MUHAMMAD, MURTALA |
collection | PubMed |
description | Streptococcus iniae is a pathogenic and zoonotic bacteria that impacted high mortality to many fish species as well as capable of causing serious disease to humans. Alanine racemase (Alr, EC 5.1.1.1) is a pyridoxal-5’-phosphate (PLP)-containing homodimeric enzyme that catalyzes the racemization of L-alanine and D-alanine. In this study, we purified alanine racemase from S. iniae that was isolated from an infected Chinese sturgeon (Acipenser sinensis), as well as determined its biochemical characteristics and inhibitors. The alr gene has an open reading frame (ORF) of 1107 bp, encoding a protein of 369 amino acids, which has a molecular mass of 40 kDa. The enzyme has optimal activity at a temperature of 35°C and a pH of 9.5. It belongs to the PLP-dependent enzymes family and is highly specific to L-alanine. S. iniae Alr (SiAlr) could be inhibited by some metal ions, hydroxylamine and dithiothreitol (DTT). The kinetic parameters K(m) and V(max) of the enzyme were 33.11 mM, 2426 units/mg for L-alanine, and 14.36 mM, 963.6 units/mg for D-alanine. Finally, the 50% inhibitory concentrations (IC(50)) values and antibiotic activity of two alanine racemase inhibitors (homogentisic acid and hydroquinone), were determined and found to be effective against both Gram-positive and Gram-negative bacteria employed in this study. |
format | Online Article Text |
id | pubmed-7256847 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Exeley Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-72568472020-06-03 Purification, Characterization and Inhibition of Alanine Racemase from a Pathogenic Strain of Streptococcus iniae MUHAMMAD, MURTALA LI, YANGYANG GONG, SIYU SHI, YANMIN JU, JIANSONG ZHAO, BAOHUA LIU, DONG Pol J Microbiol Microbiology Streptococcus iniae is a pathogenic and zoonotic bacteria that impacted high mortality to many fish species as well as capable of causing serious disease to humans. Alanine racemase (Alr, EC 5.1.1.1) is a pyridoxal-5’-phosphate (PLP)-containing homodimeric enzyme that catalyzes the racemization of L-alanine and D-alanine. In this study, we purified alanine racemase from S. iniae that was isolated from an infected Chinese sturgeon (Acipenser sinensis), as well as determined its biochemical characteristics and inhibitors. The alr gene has an open reading frame (ORF) of 1107 bp, encoding a protein of 369 amino acids, which has a molecular mass of 40 kDa. The enzyme has optimal activity at a temperature of 35°C and a pH of 9.5. It belongs to the PLP-dependent enzymes family and is highly specific to L-alanine. S. iniae Alr (SiAlr) could be inhibited by some metal ions, hydroxylamine and dithiothreitol (DTT). The kinetic parameters K(m) and V(max) of the enzyme were 33.11 mM, 2426 units/mg for L-alanine, and 14.36 mM, 963.6 units/mg for D-alanine. Finally, the 50% inhibitory concentrations (IC(50)) values and antibiotic activity of two alanine racemase inhibitors (homogentisic acid and hydroquinone), were determined and found to be effective against both Gram-positive and Gram-negative bacteria employed in this study. Exeley Inc. 2019-09 2019-09-03 /pmc/articles/PMC7256847/ /pubmed/31880879 http://dx.doi.org/10.33073/pjm-2019-036 Text en © 2019 Murtala Muhammad et al. https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License (https://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Microbiology MUHAMMAD, MURTALA LI, YANGYANG GONG, SIYU SHI, YANMIN JU, JIANSONG ZHAO, BAOHUA LIU, DONG Purification, Characterization and Inhibition of Alanine Racemase from a Pathogenic Strain of Streptococcus iniae |
title | Purification, Characterization and Inhibition of Alanine Racemase from a Pathogenic Strain of Streptococcus iniae |
title_full | Purification, Characterization and Inhibition of Alanine Racemase from a Pathogenic Strain of Streptococcus iniae |
title_fullStr | Purification, Characterization and Inhibition of Alanine Racemase from a Pathogenic Strain of Streptococcus iniae |
title_full_unstemmed | Purification, Characterization and Inhibition of Alanine Racemase from a Pathogenic Strain of Streptococcus iniae |
title_short | Purification, Characterization and Inhibition of Alanine Racemase from a Pathogenic Strain of Streptococcus iniae |
title_sort | purification, characterization and inhibition of alanine racemase from a pathogenic strain of streptococcus iniae |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7256847/ https://www.ncbi.nlm.nih.gov/pubmed/31880879 http://dx.doi.org/10.33073/pjm-2019-036 |
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