Schleinitz Kathleen M, Schmeling Sirko, Jehmlich Nico, von Bergen Martin, Harms Hauke, Kleinsteuber Sabine, Vogt Carsten, Fuchs Georg
Helmholtz Centre for Environmental Research, Department of Environmental Microbiology, Leipzig, Germany.
Appl Environ Microbiol. 2009 Jun;75(12):3912-9. doi: 10.1128/AEM.01525-08. Epub 2009 Apr 17.
Information on anaerobic phenol metabolism by physiological groups of bacteria other than nitrate reducers is scarce. We investigated phenol degradation in the strictly anaerobic iron-reducing deltaproteobacterium Geobacter metallireducens GS-15 using metabolite, transcriptome, proteome, and enzyme analyses. The results showed that the initial steps of phenol degradation are accomplished by phenylphosphate synthase (encoded by pps genes) and phenylphosphate carboxylase (encoded by ppc genes) as known from Thauera aromatica, but they also revealed some distinct differences. The pps-ppc gene cluster identified in the genome is functional, as shown by transcription analysis. In contrast to T. aromatica, transcription of the pps- and ppc-like genes was induced not only during growth on phenol, but also during growth on benzoate. In contrast, proteins were detected only during growth on phenol, suggesting the existence of a posttranscriptional regulation mechanism for these initial steps. Phenylphosphate synthase and phenylphosphate carboxylase activities were detected in cell extracts. The carboxylase does not catalyze an isotope exchange reaction between (14)CO(2) and 4-hydroxybenzoate, which is characteristic of the T. aromatica enzyme. Whereas the enzyme of T. aromatica is encoded by ppcABCD, the pps-ppc gene cluster of G. metallireducens contains only a ppcB homologue. Nearby, but oriented in the opposite direction, is a ppcD homologue that is transcribed during growth on phenol. Genome analysis did not reveal obvious homologues of ppcA and ppcC, leaving open the question of whether these genes are dispensable for phenylphosphate carboxylase activity in G. metallireducens or are quite different from the Thauera counterparts and located elsewhere in the genome.
除了硝酸盐还原菌外,关于其他生理菌群厌氧代谢苯酚的信息很少。我们使用代谢物、转录组、蛋白质组和酶分析方法,研究了严格厌氧的铁还原δ-变形菌嗜金属地杆菌Geobacter metallireducens GS-15中苯酚的降解情况。结果表明,苯酚降解的初始步骤是由已知的来自芳香陶厄氏菌(Thauera aromatica)的苯基磷酸合酶(由pps基因编码)和苯基磷酸羧化酶(由ppc基因编码)完成的,但也揭示了一些明显的差异。基因组中鉴定出的pps-ppc基因簇是有功能的,转录分析表明了这一点。与芳香陶厄氏菌不同,pps和ppc样基因的转录不仅在苯酚生长期间被诱导,而且在苯甲酸生长期间也被诱导。相反,蛋白质仅在苯酚生长期间被检测到,这表明这些初始步骤存在转录后调控机制。在细胞提取物中检测到了苯基磷酸合酶和苯基磷酸羧化酶活性。该羧化酶不催化(14)CO(2)与4-羟基苯甲酸之间的同位素交换反应,这是芳香陶厄氏菌酶的特征。芳香陶厄氏菌的酶由ppcABCD编码,而嗜金属地杆菌的pps-ppc基因簇仅包含一个ppcB同源物。在其附近但方向相反的是一个ppcD同源物,它在苯酚生长期间被转录。基因组分析未发现ppcA和ppcC的明显同源物,这使得嗜金属地杆菌中这些基因对于苯基磷酸羧化酶活性是否可有可无,或者与芳香陶厄氏菌的对应基因有很大不同且位于基因组其他位置的问题仍未解决。