Biochemical Characterization and Phylogenetic Analysis of the Virulence Factor Lysine Decarboxylase From Vibrio vulnificus
Cadaverine is produced in organisms from the amino acid lysine in a decarboxylation reaction catalyzed by lysine decarboxylase (EC 4.1.1.18). The inducible lysine decarboxylase CadA plays a vital role in acid stress response for enteric bacteria. Vibrio vulnificus is an extremely virulent human path...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-12-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2018.03082/full |
| _version_ | 1817999115913527296 |
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| author | Lifen Han Lifen Han Jinjin Yuan Jinjin Yuan Xiulan Ao Xiulan Ao Shujin Lin Shujin Lin Xiao Han Hanhui Ye Hanhui Ye |
| author_facet | Lifen Han Lifen Han Jinjin Yuan Jinjin Yuan Xiulan Ao Xiulan Ao Shujin Lin Shujin Lin Xiao Han Hanhui Ye Hanhui Ye |
| author_sort | Lifen Han |
| collection | DOAJ |
| description | Cadaverine is produced in organisms from the amino acid lysine in a decarboxylation reaction catalyzed by lysine decarboxylase (EC 4.1.1.18). The inducible lysine decarboxylase CadA plays a vital role in acid stress response for enteric bacteria. Vibrio vulnificus is an extremely virulent human pathogen causing gastroenteritis when the acid conditions that prevent survival of V. vulnificus in the stomach or small intestine are overcome. A gene encoding CadA was identified from V. vulnificus. Subsequent analyses showed that CadA from V. vulnificus (VvCadA) is a decamer with a 82-kDa subunit. Homogenous VvCadA was purified from Escherichia coli and used for lysine decarboxylation with an optimal pH of 6.0 and optimal temperature of 37°C. The apparent Vmax and Km for lysine were 9.45 ± 0.24 μM/min and 0.45 ± 0.05 mM, respectively. Mutation analysis suggested that the amino-acid-binding pyridoxal phosphate, the cofactor of the enzyme, plays a vital role in the reaction. Mutation of the negatively charged residues interacting with lysine also affected the activity of the enzyme to some extent. Quantitative RT-PCR showed that expression of VvcadA was up-regulated under low pH, low salinity, and oxidative stresses. Furthermore, the concentration of cadaverine released to the cell exterior also increased under these stresses. Protein sequence similarity network (SSN) analysis indicated that lysine decarboxylases with ornithine decarboxylases and arginine decarboxylases shared a common ancestor, and that lysine decarboxylases are more conserved during evolution. Our data provide evidence for the biochemical characteristics and important roles of VvCadA under stress conditions. |
| first_indexed | 2024-04-14T03:03:48Z |
| format | Article |
| id | doaj.art-80606dba515848f6acfa9c41210a5e24 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-14T03:03:48Z |
| publishDate | 2018-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-80606dba515848f6acfa9c41210a5e242022-12-22T02:15:49ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-12-01910.3389/fmicb.2018.03082423074Biochemical Characterization and Phylogenetic Analysis of the Virulence Factor Lysine Decarboxylase From Vibrio vulnificusLifen Han0Lifen Han1Jinjin Yuan2Jinjin Yuan3Xiulan Ao4Xiulan Ao5Shujin Lin6Shujin Lin7Xiao Han8Hanhui Ye9Hanhui Ye10The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital, Fujian Medical University, Fuzhou, ChinaInfectious Diseases Hospital of Fuzhou, Fuzhou, ChinaThe United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital, Fujian Medical University, Fuzhou, ChinaInfectious Diseases Hospital of Fuzhou, Fuzhou, ChinaThe United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital, Fujian Medical University, Fuzhou, ChinaInfectious Diseases Hospital of Fuzhou, Fuzhou, ChinaThe United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital, Fujian Medical University, Fuzhou, ChinaInfectious Diseases Hospital of Fuzhou, Fuzhou, ChinaCollege of Biological Science and Engineering, Fuzhou University, Fuzhou, ChinaThe United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital, Fujian Medical University, Fuzhou, ChinaInfectious Diseases Hospital of Fuzhou, Fuzhou, ChinaCadaverine is produced in organisms from the amino acid lysine in a decarboxylation reaction catalyzed by lysine decarboxylase (EC 4.1.1.18). The inducible lysine decarboxylase CadA plays a vital role in acid stress response for enteric bacteria. Vibrio vulnificus is an extremely virulent human pathogen causing gastroenteritis when the acid conditions that prevent survival of V. vulnificus in the stomach or small intestine are overcome. A gene encoding CadA was identified from V. vulnificus. Subsequent analyses showed that CadA from V. vulnificus (VvCadA) is a decamer with a 82-kDa subunit. Homogenous VvCadA was purified from Escherichia coli and used for lysine decarboxylation with an optimal pH of 6.0 and optimal temperature of 37°C. The apparent Vmax and Km for lysine were 9.45 ± 0.24 μM/min and 0.45 ± 0.05 mM, respectively. Mutation analysis suggested that the amino-acid-binding pyridoxal phosphate, the cofactor of the enzyme, plays a vital role in the reaction. Mutation of the negatively charged residues interacting with lysine also affected the activity of the enzyme to some extent. Quantitative RT-PCR showed that expression of VvcadA was up-regulated under low pH, low salinity, and oxidative stresses. Furthermore, the concentration of cadaverine released to the cell exterior also increased under these stresses. Protein sequence similarity network (SSN) analysis indicated that lysine decarboxylases with ornithine decarboxylases and arginine decarboxylases shared a common ancestor, and that lysine decarboxylases are more conserved during evolution. Our data provide evidence for the biochemical characteristics and important roles of VvCadA under stress conditions.https://www.frontiersin.org/article/10.3389/fmicb.2018.03082/fullVibrio vulnificuslysine decarboxylasevirulence factorphylogenetic analysisbiochemical characterization |
| spellingShingle | Lifen Han Lifen Han Jinjin Yuan Jinjin Yuan Xiulan Ao Xiulan Ao Shujin Lin Shujin Lin Xiao Han Hanhui Ye Hanhui Ye Biochemical Characterization and Phylogenetic Analysis of the Virulence Factor Lysine Decarboxylase From Vibrio vulnificus Frontiers in Microbiology Vibrio vulnificus lysine decarboxylase virulence factor phylogenetic analysis biochemical characterization |
| title | Biochemical Characterization and Phylogenetic Analysis of the Virulence Factor Lysine Decarboxylase From Vibrio vulnificus |
| title_full | Biochemical Characterization and Phylogenetic Analysis of the Virulence Factor Lysine Decarboxylase From Vibrio vulnificus |
| title_fullStr | Biochemical Characterization and Phylogenetic Analysis of the Virulence Factor Lysine Decarboxylase From Vibrio vulnificus |
| title_full_unstemmed | Biochemical Characterization and Phylogenetic Analysis of the Virulence Factor Lysine Decarboxylase From Vibrio vulnificus |
| title_short | Biochemical Characterization and Phylogenetic Analysis of the Virulence Factor Lysine Decarboxylase From Vibrio vulnificus |
| title_sort | biochemical characterization and phylogenetic analysis of the virulence factor lysine decarboxylase from vibrio vulnificus |
| topic | Vibrio vulnificus lysine decarboxylase virulence factor phylogenetic analysis biochemical characterization |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2018.03082/full |
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