The Mechanism of Aniline Blue Degradation by Short-Chain Dehydrogenase (<i>SDRz</i>) in <i>Comamonas testosteroni</i>
Dye wastewater pollution, particularly from persistent and toxic polycyclic organic pollutants, such as aniline blue, poses a significant environmental challenge. Aniline blue, a triphenylmethane dye widely used in the textile, leather, paper, and pharmaceutical industries, is notoriously difficult...
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2024-11-01
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| author | Chuanzhi Zhang Yong Huang Jiaxin He Lei He Jinyuan Zhang Lijing Yu Elshan Musazade Edmund Maser Guangming Xiong Miao Xu Liquan Guo |
| author_facet | Chuanzhi Zhang Yong Huang Jiaxin He Lei He Jinyuan Zhang Lijing Yu Elshan Musazade Edmund Maser Guangming Xiong Miao Xu Liquan Guo |
| author_sort | Chuanzhi Zhang |
| collection | DOAJ |
| description | Dye wastewater pollution, particularly from persistent and toxic polycyclic organic pollutants, such as aniline blue, poses a significant environmental challenge. Aniline blue, a triphenylmethane dye widely used in the textile, leather, paper, and pharmaceutical industries, is notoriously difficult to treat owing to its complex structure and potential for bioaccumulation. In this study, we explored the capacity of <i>Comamonas testosteroni</i> (CT1) to efficiently degrade aniline blue, focusing on the underlying enzymatic mechanisms and degradation pathways. Through prokaryotic transcriptome analysis, we identified a significantly upregulated short-chain dehydrogenase (<i>SDRz</i>) gene (log<sub>2</sub>FC = 2.11, <i>p</i> < 0.05) that plays a crucial role in the degradation process. The SDRz enzyme possessed highly conserved motifs and a typical short-chain dehydrogenase structure. Functional validation using an <i>SDRz</i>-knockout strain (CT-ΔSDRz) and an <i>SDRz</i>-expressioning strains (E-SDRz) confirmed that SDRz is essential for aniline blue degradation. The knockout strain CT-ΔSDRz exhibited a 1.27-fold reduction in the degradation efficiency, compared to CT1 strain after 12 h; while the expression strain E-SDRz showed a 1.24-fold increase compared to <i>Escherichia coli</i> DH5α after 12 h. Recombinant SDRz (rSDRz) was successfully produced, showing significant enzymatic activity (1.267 ± 0.04 mmol·L<sup>−1</sup>·min<sup>−1</sup> protein), with kinetic parameters Vmax = 2.870 ± 0.0156 mmol·L⁻<sup>1</sup>·min⁻<sup>1</sup> protein and Km = 1.805 ± 0.0128 mM·mL<sup>−1</sup>. Under optimal conditions, the rSDRz achieved a degradation efficiency of 62.17% for aniline blue. Gas chromatography–mass spectrometry (GC-MS) analysis identified several intermediate metabolites in the degradation pathway, including benzeneacetaldehyde, a, a-diphenyl, 2-amino-4-methylbenzophenone, benzene, 1-dimethylamino-4-phenylmethyl, benzenesulfonic acid, methyl ester, further elucidating the biodegradation mechanism. These findings highlight SDRz as a critical enzyme in the biodegradation of aniline blue, offering valuable insights and a robust theoretical foundation for developing advanced bioremediation strategies to address dye wastewater pollution. |
| format | Article |
| id | doaj-art-fb55a1a13eba4642bd2ece1e83203861 |
| institution | OA Journals |
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| spelling | doaj-art-fb55a1a13eba4642bd2ece1e832038612025-08-20T02:04:55ZengMDPI AGMolecules1420-30492024-11-012922540510.3390/molecules29225405The Mechanism of Aniline Blue Degradation by Short-Chain Dehydrogenase (<i>SDRz</i>) in <i>Comamonas testosteroni</i>Chuanzhi Zhang0Yong Huang1Jiaxin He2Lei He3Jinyuan Zhang4Lijing Yu5Elshan Musazade6Edmund Maser7Guangming Xiong8Miao Xu9Liquan Guo10College of Life Sciences, Jilin Agricultural University, Changchun 130118, ChinaCollege of Life Sciences, Jilin Agricultural University, Changchun 130118, ChinaCollege of Life Sciences, Jilin Agricultural University, Changchun 130118, ChinaCollege of Life Sciences, Jilin Agricultural University, Changchun 130118, ChinaCollege of Life Sciences, Jilin Agricultural University, Changchun 130118, ChinaSchool of Food and Biology, Changchun Polytechnic, Changchun 130033, ChinaCollege of Life Sciences, Jilin Agricultural University, Changchun 130118, ChinaInstitute of Toxicology and Pharmacology, University Medical School Schleswig-Holstein, 24105 Kiel, GermanyInstitute of Toxicology and Pharmacology, University Medical School Schleswig-Holstein, 24105 Kiel, GermanyCollege of Life Sciences, Jilin Agricultural University, Changchun 130118, ChinaCollege of Life Sciences, Jilin Agricultural University, Changchun 130118, ChinaDye wastewater pollution, particularly from persistent and toxic polycyclic organic pollutants, such as aniline blue, poses a significant environmental challenge. Aniline blue, a triphenylmethane dye widely used in the textile, leather, paper, and pharmaceutical industries, is notoriously difficult to treat owing to its complex structure and potential for bioaccumulation. In this study, we explored the capacity of <i>Comamonas testosteroni</i> (CT1) to efficiently degrade aniline blue, focusing on the underlying enzymatic mechanisms and degradation pathways. Through prokaryotic transcriptome analysis, we identified a significantly upregulated short-chain dehydrogenase (<i>SDRz</i>) gene (log<sub>2</sub>FC = 2.11, <i>p</i> < 0.05) that plays a crucial role in the degradation process. The SDRz enzyme possessed highly conserved motifs and a typical short-chain dehydrogenase structure. Functional validation using an <i>SDRz</i>-knockout strain (CT-ΔSDRz) and an <i>SDRz</i>-expressioning strains (E-SDRz) confirmed that SDRz is essential for aniline blue degradation. The knockout strain CT-ΔSDRz exhibited a 1.27-fold reduction in the degradation efficiency, compared to CT1 strain after 12 h; while the expression strain E-SDRz showed a 1.24-fold increase compared to <i>Escherichia coli</i> DH5α after 12 h. Recombinant SDRz (rSDRz) was successfully produced, showing significant enzymatic activity (1.267 ± 0.04 mmol·L<sup>−1</sup>·min<sup>−1</sup> protein), with kinetic parameters Vmax = 2.870 ± 0.0156 mmol·L⁻<sup>1</sup>·min⁻<sup>1</sup> protein and Km = 1.805 ± 0.0128 mM·mL<sup>−1</sup>. Under optimal conditions, the rSDRz achieved a degradation efficiency of 62.17% for aniline blue. Gas chromatography–mass spectrometry (GC-MS) analysis identified several intermediate metabolites in the degradation pathway, including benzeneacetaldehyde, a, a-diphenyl, 2-amino-4-methylbenzophenone, benzene, 1-dimethylamino-4-phenylmethyl, benzenesulfonic acid, methyl ester, further elucidating the biodegradation mechanism. These findings highlight SDRz as a critical enzyme in the biodegradation of aniline blue, offering valuable insights and a robust theoretical foundation for developing advanced bioremediation strategies to address dye wastewater pollution.https://www.mdpi.com/1420-3049/29/22/5405<i>Comamonas testosteroni</i>short-chain dehydrogenaseaniline bluebiodegradation |
| spellingShingle | Chuanzhi Zhang Yong Huang Jiaxin He Lei He Jinyuan Zhang Lijing Yu Elshan Musazade Edmund Maser Guangming Xiong Miao Xu Liquan Guo The Mechanism of Aniline Blue Degradation by Short-Chain Dehydrogenase (<i>SDRz</i>) in <i>Comamonas testosteroni</i> Molecules <i>Comamonas testosteroni</i> short-chain dehydrogenase aniline blue biodegradation |
| title | The Mechanism of Aniline Blue Degradation by Short-Chain Dehydrogenase (<i>SDRz</i>) in <i>Comamonas testosteroni</i> |
| title_full | The Mechanism of Aniline Blue Degradation by Short-Chain Dehydrogenase (<i>SDRz</i>) in <i>Comamonas testosteroni</i> |
| title_fullStr | The Mechanism of Aniline Blue Degradation by Short-Chain Dehydrogenase (<i>SDRz</i>) in <i>Comamonas testosteroni</i> |
| title_full_unstemmed | The Mechanism of Aniline Blue Degradation by Short-Chain Dehydrogenase (<i>SDRz</i>) in <i>Comamonas testosteroni</i> |
| title_short | The Mechanism of Aniline Blue Degradation by Short-Chain Dehydrogenase (<i>SDRz</i>) in <i>Comamonas testosteroni</i> |
| title_sort | mechanism of aniline blue degradation by short chain dehydrogenase i sdrz i in i comamonas testosteroni i |
| topic | <i>Comamonas testosteroni</i> short-chain dehydrogenase aniline blue biodegradation |
| url | https://www.mdpi.com/1420-3049/29/22/5405 |
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