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| Microbial degradation pathway of dibutylphthalate by a typical ultramicrobacterium and the expression and purification of 4-oxalomesaconate hydratase |
| Received:August 06, 2016 |
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| KeyWord:ultramicrobacterium;phthalate esters;4-oxalomesaconate hydratase(OMH);X-ray;protein expression and purification |
| Author Name | Affiliation | E-mail | | MA Dan | College of Environmental Science and Engineering, Nankai University, Key Laboratory of Pollution Processes and Environmental Criteria(Ministry of Education), Tianjin Key Laboratory of Environmental Remediation and Pollution Control, Tianjin 300071, China | | | SUN Rui | College of Environmental Science and Engineering, Nankai University, Key Laboratory of Pollution Processes and Environmental Criteria(Ministry of Education), Tianjin Key Laboratory of Environmental Remediation and Pollution Control, Tianjin 300071, China | | | QUAN Wei | College of Environmental Science and Engineering, Nankai University, Key Laboratory of Pollution Processes and Environmental Criteria(Ministry of Education), Tianjin Key Laboratory of Environmental Remediation and Pollution Control, Tianjin 300071, China | | | WANG Ying-ying | College of Environmental Science and Engineering, Nankai University, Key Laboratory of Pollution Processes and Environmental Criteria(Ministry of Education), Tianjin Key Laboratory of Environmental Remediation and Pollution Control, Tianjin 300071, China | wangyy@nankai.edu.cn |
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| Abstract: |
| A typical ultramicrobacterium, Curvibacter sp. strain PAE-UM, capable of degrading dibutyl phthalate was isolated. It is a Gram negative strain. Flow cytometer was used to detect the bacterial growth and the HPLC-MS was applied to analyze the concentration of DBP. The result showed that this strain can degrade DBP with a degradation rate of 94% and the maximum growth rate of strain was 0.237 2 h-1. Its draft genome has been sequenced in our previous study. The complete genome sequence has been deposited in GenBank under the accession number LKCX00000000. In this study, we proposed the degradation pathway of dibutyl phthalate and the key enzymes involved based on the genome analysis. The results showed that 4-oxalomesaconate hydratase(OMH) was the key enzyme in the meta-cleavage of protocatechuic acid in PAEs degradation pathway. It is also the key enzyme in the aromatic compounds degradation pathway. It has the ability to catalyze the 4-oxalomesaconate to 4-carboxy-4-hydroxy-2-oxoadipate. The gene encoding OMH was expressed in E. coli. The protein was then purified by a variety of methods. Protein crystals with high resolution were obtained. The crystal data was then collected by X-ray diffraction experiment in Shanghai Synchrotron Radiation Facility. The data was processed by the HKL2000 software and the resolution was 2.0 Å. These results could make an important contribution to the study of OMH functional mechanism. Furthermore, the results could further clarify the molecular mechanism of the PAEs degradation by ultramicrobacteria. |
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