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The CDHB hydrolase (CbaA) was purified from strain DL-8, which can also hydrolyze 2-benzoxazolinone (BOA), 5-chloro-2-BOA, and benzamide. The specific activity of purified CbaA was 5,900 U \u00b7 mg protein\n                    <jats:sup>\u22121<\/jats:sup>\n                    for CDHB, with\n                    <jats:italic>\n                      K\n                      <jats:sub>m<\/jats:sub>\n                    <\/jats:italic>\n                    and\n                    <jats:italic>k<\/jats:italic>\n                    <jats:sub>cat<\/jats:sub>\n                    values of 0.29 mM and 8,500 s\n                    <jats:sup>\u22121<\/jats:sup>\n                    , respectively. The optimal pH for purified CbaA was 9.0, the highest activity was observed at 55\u00b0C, and the inactive metal-free enzyme could be reactivated by Mg\n                    <jats:sup>2+<\/jats:sup>\n                    , Ni\n                    <jats:sup>2+<\/jats:sup>\n                    , Ca\n                    <jats:sup>2+<\/jats:sup>\n                    , or Zn\n                    <jats:sup>2+<\/jats:sup>\n                    . Based on the results obtained for the CbaA peptide mass fingerprinting and draft genome sequence of strain DL-8,\n                    <jats:italic>cbaA<\/jats:italic>\n                    (encoding 339 amino acids) was cloned and expressed in\n                    <jats:named-content content-type=\"genus-species\">Escherichia coli<\/jats:named-content>\n                    BL21(DE3). CbaA shared 18 to 21% identity with some metal-dependent hydrolases of the PF01499 family and contained the signature metal-binding motif Q\n                    <jats:sub>127<\/jats:sub>\n                    XXXQ\n                    <jats:sub>131<\/jats:sub>\n                    XD\n                    <jats:sub>133<\/jats:sub>\n                    XXXH\n                    <jats:sub>137<\/jats:sub>\n                    . The conserved amino acid residues His288 and Glu301 served as the proton donor and acceptor.\n                    <jats:named-content content-type=\"genus-species\">E. coli<\/jats:named-content>\n                    BL21(DE3-pET-\n                    <jats:italic>cbaA<\/jats:italic>\n                    ) resting cells could transform 0.2 mM CDHB into 2A5CP. The mutant strain DL-8\u0394\n                    <jats:italic>cbaA<\/jats:italic>\n                    lost the ability to degrade CDHB but retained the ability to degrade 2A5CP, consistent with strain DL-8. These results indicated that\n                    <jats:italic>cbaA<\/jats:italic>\n                    was the key gene responsible for CDHB degradation by strain DL-8.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>IMPORTANCE<\/jats:bold>\n                    2-Benzoxazolinone (BOA) derivatives are widely used as synthetic intermediates and are also an important group of allelochemicals acting in response to tissue damage or pathogen attack in gramineous plants. However, the degradation mechanism of BOA derivatives by microorganisms is not clear. In the present study, we reported the identification of CbaA and metabolic pathway responsible for the degradation of CDHB in\n                    <jats:named-content content-type=\"genus-species\">Pigmentiphaga<\/jats:named-content>\n                    sp. DL-8. This will provide microorganism and gene resources for the bioremediation of the environmental pollution caused by BOA derivatives.\n                  <\/jats:p>","DOI":"10.1128\/aem.00532-16","type":"journal-article","created":{"date-parts":[[2016,5,7]],"date-time":"2016-05-07T07:49:59Z","timestamp":1462607399000},"page":"4169-4179","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":15,"title":["Metabolic Pathway Involved in 6-Chloro-2-Benzoxazolinone Degradation by Pigmentiphaga sp. Strain DL-8 and Identification of the Novel Metal-Dependent Hydrolase CbaA"],"prefix":"10.1128","volume":"82","author":[{"given":"Weiliang","family":"Dong","sequence":"first","affiliation":[{"name":"Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, People's Republic of China"},{"name":"State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fei","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, People's Republic of China"},{"name":"College of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang, People's Republic 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