Growth of Dehalococcoides mccartyi strain CBDB1 by reductive dehalogenation of brominated benzenes to benzene.
Growth of Dehalococcoides mccartyi strain CBDB1 by reductive dehalogenation of brominated benzenes to benzene.
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DOI：
10.1021/es3003519
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发表时间：
2012-08
影响因子：
11.4
通讯作者：
Anke Wagner;Myriel Cooper;Sara Ferdi;J. Seifert;L. Adrian
中科院分区：
文献类型：
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作者：
Anke Wagner;Myriel Cooper;Sara Ferdi;J. Seifert;L. Adrian
溴化芳族化合物用于许多不同的应用中，但也天然存在。在这里，我们证明了有机卤化物呼吸和生长Dehalococcoides mccartyi菌株CBDB 1与1，2，4-三溴苯，所有三个二溴苯同系物和一溴苯。所有溴苯完全脱卤为苯。每摩尔释放的溴化物的生长产量在1.8 × 10（14）和2.8 × 10（14）个细胞之间。此外，建立了一种新设计的高通量基于甲基紫精的光度微量滴定板测定法，以测定以溴化芳香族化合物为电子受体的CBDB 1菌株静息细胞测定中还原性脱卤酶的活性。在1，2，4-三溴苯上培养菌株CBDB 1后，计算出2.8-13.2 nkat/mg总细胞蛋白（0.16-0.8单位/mg总细胞蛋白）的活性。用菌株CBDB 1的全细胞提取物进行的质谱分析和活性测定给出了强有力的证据，即四到六种还原性脱卤酶参与所有测试的溴化苯的脱卤，包括还原性脱卤酶CbdbA 80和CbrA。
Brominated aromatics are used in many different applications but occur also naturally. Here, we demonstrate organohalide respiration and growth of Dehalococcoides mccartyi strain CBDB1 with 1,2,4-tribromobenzene, all three dibrominated benzene congeners and monobromobenzene. All bromobenzenes were fully dehalogenated to benzene. Growth yields were between 1.8 × 10(14) and 2.8 × 10(14) cells per mol of bromide released. Furthermore, a newly designed high-throughput methyl viologen-based photometric microtiter plate assay was established to determine the activity of the reductive dehalogenases in resting cell assays of strain CBDB1 with brominated aromatics as electron acceptors. Activities of 2.8-13.2 nkat per mg total cell protein (0.16-0.8 units per mg total cell protein) were calculated after cultivation of strain CBDB1 on 1,2,4-tribromobenzene. Mass spectrometric analyses and activity assays with whole cell extracts of strain CBDB1 gave strong evidence that four to six reductive dehalogenases were involved in the dehalogenation of all tested brominated benzenes, including the reductive dehalogenases CbdbA80 and CbrA.