Schleheck David, Cook Alasdair M
Department of Biology, The University of Konstanz, 78457 Konstanz, Germany.
Arch Microbiol. 2005 Sep;183(6):369-77. doi: 10.1007/s00203-005-0002-7. Epub 2005 Oct 13.
Parvibaculum lavamentivorans (T) DS-1, an aerobic, heterotrophic bacterium, requires a biofilm on a solid surface (e.g. glass particles) when utilizing commercial linear alkylbenzenesulfonate surfactant (LAS; 20 congeners) for growth. Catabolism involves the undefined 'omega-oxygenation' and beta-oxidation of the LAS side chain, and the organism excretes sulfophenyl carboxylates (SPC) quantitatively. A 3.5-l fermenter was developed which allowed gram-quantities of LAS-grown cells to be grown and harvested from medium with glass particles as the solid support. The catabolism of LAS was dominant: in diauxie experiments with acetate as second carbon source, LAS was utilized first. The biofilm-encoated LAS-grown cells were unsuitable for metabolic work in vitro because cell suspensions clumped and were not disrupted effectively, but the degradative enzymes were found to be expressed constitutively in acetate-grown cells, which formed no biofilm. LAS-dependent oxygen uptake was measured in acetate-grown cells at about 0.6 mkat (kg protein)(-1), but not in extracts of cells. Whole cells converted LAS to SPC in the presence of molecular oxygen only, and the reaction could be saturably inhibited by metyrapone, which acts on e.g. cytochromes P450 (CYP). However, despite the presence of CYP153-like sequences in the genome of strain DS-1(T), the difference spectra did not support the presence of a CYP in crude extracts, and the nature of the LAS-oxygenase remains unclear.
洗烷微小杆菌(模式菌株)DS-1是一种需氧异养细菌,在利用商业直链烷基苯磺酸盐表面活性剂(LAS;20种同系物)进行生长时,需要在固体表面(如玻璃颗粒)形成生物膜。其分解代谢涉及LAS侧链未明确的“ω-氧化”和β-氧化,并且该生物体定量分泌磺苯基羧酸盐(SPC)。开发了一种3.5升发酵罐,可在以玻璃颗粒作为固体支持物的培养基中培养并收获克数量的在LAS上生长的细胞。LAS的分解代谢占主导:在以乙酸盐作为第二碳源的双相生长实验中,LAS首先被利用。被生物膜包裹的在LAS上生长的细胞不适合用于体外代谢研究,因为细胞悬液会聚集且不能有效分散,但发现降解酶在不形成生物膜的乙酸盐生长细胞中组成性表达。在乙酸盐生长细胞中测得LAS依赖的氧摄取约为0.6 mkat(千克蛋白质)-1,但在细胞提取物中未测得。完整细胞仅在分子氧存在的情况下将LAS转化为SPC,并且该反应可被甲吡酮饱和抑制,甲吡酮作用于例如细胞色素P450(CYP)。然而,尽管DS-1(T)菌株的基因组中存在CYP153样序列,但差光谱不支持在粗提取物中存在CYP,LAS加氧酶的性质仍不清楚。