Degradation of 2,4,6-Trinitrotoluene (TNT): Involvement of Protocatechuate 3,4-Dioxygenase (P34O) in <em>Buttiauxella</em> sp. S19-1
Extensive use and disposal of 2,4,6-trinitrotoluene (TNT), a primary constituent of explosives, pollutes the environment and causes severe damage to human health. Complete mineralization of TNT via bacterial degradation has recently gained research interest as an effective method for the restoration...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-09-01
|
| Series: | Toxics |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2305-6304/9/10/231 |
| _version_ | 1827678556317024256 |
|---|---|
| author | Miao Xu Dong Liu Ping Sun Yunuo Li Ming Wu Wencong Liu Edmund Maser Guangming Xiong Liquan Guo |
| author_facet | Miao Xu Dong Liu Ping Sun Yunuo Li Ming Wu Wencong Liu Edmund Maser Guangming Xiong Liquan Guo |
| author_sort | Miao Xu |
| collection | DOAJ |
| description | Extensive use and disposal of 2,4,6-trinitrotoluene (TNT), a primary constituent of explosives, pollutes the environment and causes severe damage to human health. Complete mineralization of TNT via bacterial degradation has recently gained research interest as an effective method for the restoration of contaminated sites. Here, screening for TNT degradation by six selected bacteria revealed that <i>Buttiauxella</i> sp. S19-1, possesses the strongest degrading ability. Moreover, <i>BuP34O</i> (a gene encoding for protocatechuate 3,4-dioxygenase—P34O, a key enzyme in the β-ketoadipate pathway) was upregulated during TNT degradation. A knockout of <i>BuP34O</i> in S19-1 to generate S-M1 mutant strain caused a marked reduction in TNT degradation efficiency compared to S19-1. Additionally, the EM1 mutant strain (<i>Escherichia coli</i> DH5α transfected with <i>BuP34O</i>) showed higher degradation efficiency than DH5α. Gas chromatography mass spectrometry (GC-MS) analysis of TNT degradation by S19-1 revealed 4-amino-2,6-dinitrotolune (ADNT) as the intermediate metabolite of TNT. Furthermore, the recombinant protein P34O (rP34O) expressed the activity of 2.46 µmol/min·mg. Our findings present the first report on the involvement of P34O in bacterial degradation of TNT and its metabolites, suggesting that P34O could catalyze downstream reactions in the TNT degradation pathway. In addition, the TNT-degrading ability of S19-1, a Gram-negative marine-derived bacterium, presents enormous potential for restoration of TNT-contaminated seas. |
| first_indexed | 2024-03-10T06:10:48Z |
| format | Article |
| id | doaj.art-ccfcebf62c66400ba6b2fe1d12fd1f38 |
| institution | Directory Open Access Journal |
| issn | 2305-6304 |
| language | English |
| last_indexed | 2024-03-10T06:10:48Z |
| publishDate | 2021-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxics |
| spelling | doaj.art-ccfcebf62c66400ba6b2fe1d12fd1f382023-11-22T20:12:07ZengMDPI AGToxics2305-63042021-09-0191023110.3390/toxics9100231Degradation of 2,4,6-Trinitrotoluene (TNT): Involvement of Protocatechuate 3,4-Dioxygenase (P34O) in <em>Buttiauxella</em> sp. S19-1Miao Xu0Dong Liu1Ping Sun2Yunuo Li3Ming Wu4Wencong Liu5Edmund Maser6Guangming Xiong7Liquan Guo8Key Laboratory of Straw Biology and Higher Value Application, The Ministry of Education, College of Life Science, Jilin Agricultural University, Changchun 130118, ChinaGrain College, Jilin Busyness and Technology College, Changchun 130507, ChinaKey Laboratory of Straw Biology and Higher Value Application, The Ministry of Education, College of Life Science, Jilin Agricultural University, Changchun 130118, ChinaKey Laboratory of Straw Biology and Higher Value Application, The Ministry of Education, College of Life Science, Jilin Agricultural University, Changchun 130118, ChinaKey Laboratory of Straw Biology and Higher Value Application, The Ministry of Education, College of Life Science, Jilin Agricultural University, Changchun 130118, ChinaCollege of Resources and Environment, Jilin Agricultural University, Changchun 130118, ChinaInstitute of Toxicology and Pharmacology, University Medical School Schleswig-Holstein, 24105 Kiel, GermanyKey Laboratory of Straw Biology and Higher Value Application, The Ministry of Education, College of Life Science, Jilin Agricultural University, Changchun 130118, ChinaKey Laboratory of Straw Biology and Higher Value Application, The Ministry of Education, College of Life Science, Jilin Agricultural University, Changchun 130118, ChinaExtensive use and disposal of 2,4,6-trinitrotoluene (TNT), a primary constituent of explosives, pollutes the environment and causes severe damage to human health. Complete mineralization of TNT via bacterial degradation has recently gained research interest as an effective method for the restoration of contaminated sites. Here, screening for TNT degradation by six selected bacteria revealed that <i>Buttiauxella</i> sp. S19-1, possesses the strongest degrading ability. Moreover, <i>BuP34O</i> (a gene encoding for protocatechuate 3,4-dioxygenase—P34O, a key enzyme in the β-ketoadipate pathway) was upregulated during TNT degradation. A knockout of <i>BuP34O</i> in S19-1 to generate S-M1 mutant strain caused a marked reduction in TNT degradation efficiency compared to S19-1. Additionally, the EM1 mutant strain (<i>Escherichia coli</i> DH5α transfected with <i>BuP34O</i>) showed higher degradation efficiency than DH5α. Gas chromatography mass spectrometry (GC-MS) analysis of TNT degradation by S19-1 revealed 4-amino-2,6-dinitrotolune (ADNT) as the intermediate metabolite of TNT. Furthermore, the recombinant protein P34O (rP34O) expressed the activity of 2.46 µmol/min·mg. Our findings present the first report on the involvement of P34O in bacterial degradation of TNT and its metabolites, suggesting that P34O could catalyze downstream reactions in the TNT degradation pathway. In addition, the TNT-degrading ability of S19-1, a Gram-negative marine-derived bacterium, presents enormous potential for restoration of TNT-contaminated seas.https://www.mdpi.com/2305-6304/9/10/2312,4,6-trinitrotoluene (TNT)4-amino-2,6-dinitrotolunes (ADNT)<i>Buttiauxella</i> sp. S19-1protocatechuate 3,4-dioxygenase (P34O)degradationgenetic manipulation |
| spellingShingle | Miao Xu Dong Liu Ping Sun Yunuo Li Ming Wu Wencong Liu Edmund Maser Guangming Xiong Liquan Guo Degradation of 2,4,6-Trinitrotoluene (TNT): Involvement of Protocatechuate 3,4-Dioxygenase (P34O) in <em>Buttiauxella</em> sp. S19-1 Toxics 2,4,6-trinitrotoluene (TNT) 4-amino-2,6-dinitrotolunes (ADNT) <i>Buttiauxella</i> sp. S19-1 protocatechuate 3,4-dioxygenase (P34O) degradation genetic manipulation |
| title | Degradation of 2,4,6-Trinitrotoluene (TNT): Involvement of Protocatechuate 3,4-Dioxygenase (P34O) in <em>Buttiauxella</em> sp. S19-1 |
| title_full | Degradation of 2,4,6-Trinitrotoluene (TNT): Involvement of Protocatechuate 3,4-Dioxygenase (P34O) in <em>Buttiauxella</em> sp. S19-1 |
| title_fullStr | Degradation of 2,4,6-Trinitrotoluene (TNT): Involvement of Protocatechuate 3,4-Dioxygenase (P34O) in <em>Buttiauxella</em> sp. S19-1 |
| title_full_unstemmed | Degradation of 2,4,6-Trinitrotoluene (TNT): Involvement of Protocatechuate 3,4-Dioxygenase (P34O) in <em>Buttiauxella</em> sp. S19-1 |
| title_short | Degradation of 2,4,6-Trinitrotoluene (TNT): Involvement of Protocatechuate 3,4-Dioxygenase (P34O) in <em>Buttiauxella</em> sp. S19-1 |
| title_sort | degradation of 2 4 6 trinitrotoluene tnt involvement of protocatechuate 3 4 dioxygenase p34o in em buttiauxella em sp s19 1 |
| topic | 2,4,6-trinitrotoluene (TNT) 4-amino-2,6-dinitrotolunes (ADNT) <i>Buttiauxella</i> sp. S19-1 protocatechuate 3,4-dioxygenase (P34O) degradation genetic manipulation |
| url | https://www.mdpi.com/2305-6304/9/10/231 |
| work_keys_str_mv | AT miaoxu degradationof246trinitrotoluenetntinvolvementofprotocatechuate34dioxygenasep34oinembuttiauxellaemsps191 AT dongliu degradationof246trinitrotoluenetntinvolvementofprotocatechuate34dioxygenasep34oinembuttiauxellaemsps191 AT pingsun degradationof246trinitrotoluenetntinvolvementofprotocatechuate34dioxygenasep34oinembuttiauxellaemsps191 AT yunuoli degradationof246trinitrotoluenetntinvolvementofprotocatechuate34dioxygenasep34oinembuttiauxellaemsps191 AT mingwu degradationof246trinitrotoluenetntinvolvementofprotocatechuate34dioxygenasep34oinembuttiauxellaemsps191 AT wencongliu degradationof246trinitrotoluenetntinvolvementofprotocatechuate34dioxygenasep34oinembuttiauxellaemsps191 AT edmundmaser degradationof246trinitrotoluenetntinvolvementofprotocatechuate34dioxygenasep34oinembuttiauxellaemsps191 AT guangmingxiong degradationof246trinitrotoluenetntinvolvementofprotocatechuate34dioxygenasep34oinembuttiauxellaemsps191 AT liquanguo degradationof246trinitrotoluenetntinvolvementofprotocatechuate34dioxygenasep34oinembuttiauxellaemsps191 |