O'Loughlin Sheryl Naomi, Graham Robert L J, McMullan Geoff, Ternan Nigel G
School of Biomedical Sciences, University of Ulster, Coleraine, UK.
FEMS Microbiol Lett. 2006 Aug;261(1):133-40. doi: 10.1111/j.1574-6968.2006.00344.x.
A strain of Agromyces fucosus, designated Vs2, metabolized a range of organophosphonate compounds as sole phosphorus sources for growth and metabolized phosphonoacetate as a sole carbon, energy and phosphorus source for growth. With phosphonoacetate as the sole phosphorus source and a pyruvate carbon source, transient phosphate release to the medium was observed, in contrast to cultures grown with glucose and phosphonoacetate, where no phosphate release to the medium was observed. Carbon catabolite repression, specifically by means of inducer exclusion of phosphonoacetate, was proposed as the mechanism responsible, and phosphonoacetate hydrolase enzyme assays carried out on cell extracts confirmed that induced phosphonoacetate hydrolase activities were indeed higher in cells grown on pyruvate with phosphonoacetate as sole phosphorus source. This phenomenon has not previously been demonstrated in vivo, and must represent a significant metabolic control of organophosphonate metabolism. The catabolite repression phenomenon was also evident when A. fucosus grew on 2-aminoethylphosphonate as sole phosphorus source, allowing demonstration of a third mode of control for biodegradation of this compound. Excision of stained zymogram gel pieces, followed by tryptic digestion and mass spectrometric analysis, allowed the identification of phosphonoacetate hydrolase-derived peptides.
一株被命名为Vs2的岩岸土壤杆菌(Agromyces fucosus)能够将一系列有机膦酸盐化合物作为唯一的磷源进行代谢,并能将膦酰基乙酸作为唯一的碳源、能源和磷源进行代谢。以膦酰基乙酸作为唯一的磷源和丙酮酸作为碳源时,观察到向培养基中短暂释放了磷酸盐,这与以葡萄糖和膦酰基乙酸培养的培养物形成对比,后者未观察到向培养基中释放磷酸盐。提出碳分解代谢物阻遏,特别是通过膦酰基乙酸的诱导物排除作用,作为其作用机制,并且对细胞提取物进行的膦酰基乙酸水解酶活性测定证实,在以膦酰基乙酸作为唯一磷源的丙酮酸培养基上生长的细胞中,诱导的膦酰基乙酸水解酶活性确实更高。这种现象以前尚未在体内得到证实,并且必定代表了对有机膦酸盐代谢的一种重要的代谢调控。当岩岸土壤杆菌以2-氨基乙基膦酸盐作为唯一磷源生长时,分解代谢物阻遏现象也很明显,这使得能够证明该化合物生物降解的第三种调控模式。切除染色的酶谱凝胶条带,然后进行胰蛋白酶消化和质谱分析,从而鉴定出膦酰基乙酸水解酶衍生的肽段。