Molecular Insight into the Acryloyl-CoA Hydration by AcuH for Acrylate Detoxification in Dimethylsulfoniopropionate-Catabolizing Bacteria
Microbial cleavage of dimethylsulfoniopropionate (DMSP) producing dimethyl sulfide (DMS) and acrylate is an important step in global sulfur cycling. Acrylate is toxic for cells, and thus should be metabolized effectively for detoxification. There are two proposed pathways for acrylate metabolism in...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2017-10-01
|
| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/article/10.3389/fmicb.2017.02034/full |
| _version_ | 1811255104802652160 |
|---|---|
| author | Hai-Yan Cao Peng Wang Fei Xu Ping-Yi Li Bin-Bin Xie Qi-Long Qin Yu-Zhong Zhang Yu-Zhong Zhang Chun-Yang Li Xiu-Lan Chen |
| author_facet | Hai-Yan Cao Peng Wang Fei Xu Ping-Yi Li Bin-Bin Xie Qi-Long Qin Yu-Zhong Zhang Yu-Zhong Zhang Chun-Yang Li Xiu-Lan Chen |
| author_sort | Hai-Yan Cao |
| collection | DOAJ |
| description | Microbial cleavage of dimethylsulfoniopropionate (DMSP) producing dimethyl sulfide (DMS) and acrylate is an important step in global sulfur cycling. Acrylate is toxic for cells, and thus should be metabolized effectively for detoxification. There are two proposed pathways for acrylate metabolism in DMSP-catabolizing bacteria, the AcuN-AcuK pathway and the PrpE-AcuI pathway. AcuH is an acryloyl-CoA hydratase in DMSP-catabolizing bacteria and can catalyze the hydration of toxic acryloyl-CoA to produce 3-hydroxypropionyl-CoA (3-HP-CoA) in both the AcuN-AcuK pathway and the side path of the PrpE-AcuI pathway. However, the structure and catalytic mechanism of AcuH remain unknown. Here, we cloned a putative acuH gene from Roseovarius nubinhibens ISM, a typical DMSP-catabolizing bacterium, and expressed it (RdAcuH) in Escherichia coli. The activity of RdAcuH toward acryloyl-CoA was detected by liquid chromatography-mass spectrometry (LC-MS), which suggests that RdAcuH is a functional acryloyl-CoA hydratase. Then we solved the crystal structure of RdAcuH. Each asymmetric unit in the crystal of RdAcuH contains a dimer of trimers and each RdAcuH monomer contains an N-terminal domain (NTD) and a C-terminal domain (CTD). There are three active centers in each trimer and each active center is located between the NTD of a subunit and the CTD of the neighboring subunit. Site-directed mutagenesis analysis indicates that two highly conserved glutamates, Glu112 and Glu132, in the active center are essential for catalysis. Based on our results and previous research, we analyzed the catalytic mechanism of AcuH to hydrate acryloyl-CoA, in which Glu132 acts as the catalytic base. This study sheds light on the mechanism of acrylate detoxification in DMSP-catabolizing bacteria. |
| first_indexed | 2024-04-12T17:18:52Z |
| format | Article |
| id | doaj.art-43a86d27e68049e594d05c54497ab116 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-12T17:18:52Z |
| publishDate | 2017-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-43a86d27e68049e594d05c54497ab1162022-12-22T03:23:32ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2017-10-01810.3389/fmicb.2017.02034296335Molecular Insight into the Acryloyl-CoA Hydration by AcuH for Acrylate Detoxification in Dimethylsulfoniopropionate-Catabolizing BacteriaHai-Yan Cao0Peng Wang1Fei Xu2Ping-Yi Li3Bin-Bin Xie4Qi-Long Qin5Yu-Zhong Zhang6Yu-Zhong Zhang7Chun-Yang Li8Xiu-Lan Chen9State Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Institute of Marine Science and Technology, Shandong University, Jinan, ChinaState Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Institute of Marine Science and Technology, Shandong University, Jinan, ChinaState Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Institute of Marine Science and Technology, Shandong University, Jinan, ChinaState Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Institute of Marine Science and Technology, Shandong University, Jinan, ChinaState Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Institute of Marine Science and Technology, Shandong University, Jinan, ChinaState Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Institute of Marine Science and Technology, Shandong University, Jinan, ChinaState Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Institute of Marine Science and Technology, Shandong University, Jinan, ChinaLaboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, ChinaState Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Institute of Marine Science and Technology, Shandong University, Jinan, ChinaState Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Institute of Marine Science and Technology, Shandong University, Jinan, ChinaMicrobial cleavage of dimethylsulfoniopropionate (DMSP) producing dimethyl sulfide (DMS) and acrylate is an important step in global sulfur cycling. Acrylate is toxic for cells, and thus should be metabolized effectively for detoxification. There are two proposed pathways for acrylate metabolism in DMSP-catabolizing bacteria, the AcuN-AcuK pathway and the PrpE-AcuI pathway. AcuH is an acryloyl-CoA hydratase in DMSP-catabolizing bacteria and can catalyze the hydration of toxic acryloyl-CoA to produce 3-hydroxypropionyl-CoA (3-HP-CoA) in both the AcuN-AcuK pathway and the side path of the PrpE-AcuI pathway. However, the structure and catalytic mechanism of AcuH remain unknown. Here, we cloned a putative acuH gene from Roseovarius nubinhibens ISM, a typical DMSP-catabolizing bacterium, and expressed it (RdAcuH) in Escherichia coli. The activity of RdAcuH toward acryloyl-CoA was detected by liquid chromatography-mass spectrometry (LC-MS), which suggests that RdAcuH is a functional acryloyl-CoA hydratase. Then we solved the crystal structure of RdAcuH. Each asymmetric unit in the crystal of RdAcuH contains a dimer of trimers and each RdAcuH monomer contains an N-terminal domain (NTD) and a C-terminal domain (CTD). There are three active centers in each trimer and each active center is located between the NTD of a subunit and the CTD of the neighboring subunit. Site-directed mutagenesis analysis indicates that two highly conserved glutamates, Glu112 and Glu132, in the active center are essential for catalysis. Based on our results and previous research, we analyzed the catalytic mechanism of AcuH to hydrate acryloyl-CoA, in which Glu132 acts as the catalytic base. This study sheds light on the mechanism of acrylate detoxification in DMSP-catabolizing bacteria.http://journal.frontiersin.org/article/10.3389/fmicb.2017.02034/fullDMSPacrylatedetoxificationacryloyl-CoA hydratasecatalytic mechanism |
| spellingShingle | Hai-Yan Cao Peng Wang Fei Xu Ping-Yi Li Bin-Bin Xie Qi-Long Qin Yu-Zhong Zhang Yu-Zhong Zhang Chun-Yang Li Xiu-Lan Chen Molecular Insight into the Acryloyl-CoA Hydration by AcuH for Acrylate Detoxification in Dimethylsulfoniopropionate-Catabolizing Bacteria Frontiers in Microbiology DMSP acrylate detoxification acryloyl-CoA hydratase catalytic mechanism |
| title | Molecular Insight into the Acryloyl-CoA Hydration by AcuH for Acrylate Detoxification in Dimethylsulfoniopropionate-Catabolizing Bacteria |
| title_full | Molecular Insight into the Acryloyl-CoA Hydration by AcuH for Acrylate Detoxification in Dimethylsulfoniopropionate-Catabolizing Bacteria |
| title_fullStr | Molecular Insight into the Acryloyl-CoA Hydration by AcuH for Acrylate Detoxification in Dimethylsulfoniopropionate-Catabolizing Bacteria |
| title_full_unstemmed | Molecular Insight into the Acryloyl-CoA Hydration by AcuH for Acrylate Detoxification in Dimethylsulfoniopropionate-Catabolizing Bacteria |
| title_short | Molecular Insight into the Acryloyl-CoA Hydration by AcuH for Acrylate Detoxification in Dimethylsulfoniopropionate-Catabolizing Bacteria |
| title_sort | molecular insight into the acryloyl coa hydration by acuh for acrylate detoxification in dimethylsulfoniopropionate catabolizing bacteria |
| topic | DMSP acrylate detoxification acryloyl-CoA hydratase catalytic mechanism |
| url | http://journal.frontiersin.org/article/10.3389/fmicb.2017.02034/full |
| work_keys_str_mv | AT haiyancao molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria AT pengwang molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria AT feixu molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria AT pingyili molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria AT binbinxie molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria AT qilongqin molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria AT yuzhongzhang molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria AT yuzhongzhang molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria AT chunyangli molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria AT xiulanchen molecularinsightintotheacryloylcoahydrationbyacuhforacrylatedetoxificationindimethylsulfoniopropionatecatabolizingbacteria |