Whitman B E, Lueking D R, Mihelcic J R
Department of GeoEnvironmental Science and Engineering, Wilkes University, Wilkes-Barre, PA 18766, USA.
Can J Microbiol. 1998 Nov;44(11):1086-93. doi: 10.1139/cjm-44-11-1086.
The uptake of naphthalene has been investigated in the metabolizing cells of Pseudomonas fluorescens utilizing [1-14C]naphthalene. The uptake displayed an affinity constant (Kt) of 11 microM and a maximal velocity (Vmax) of 17 nmol.h-1.mg-1 cellular dry weight. Naphthalene uptake was not observed in a mutant strain, TG-5, which was unable to utilize naphthalene as a sole source of carbon for growth. Uptake was significantly inhibited (approximately 90%) by the presence of growth-inhibiting levels of either azide or 2,4-dinitrophenol and was sensitive to the presence of structural analogues of naphthalene. The intracellular levels of ATP were not significantly reduced by the presence of either azide or 2,4-dinitrophenol. The presence of alpha-naphthol was found to noncompetitively inhibit naphthalene uptake, displaying a Ki of 0.041 microM. It is concluded that the first step in the utilization of naphthalene by Pseudomonas fluorescens is its transport into the cell by a specific energy-linked transport system.
利用[1-14C]萘对荧光假单胞菌的代谢细胞中萘的摄取进行了研究。摄取显示出11微摩尔的亲和常数(Kt)和17纳摩尔·小时-1·毫克-1细胞干重的最大速度(Vmax)。在突变株TG-5中未观察到萘的摄取,该突变株无法利用萘作为唯一碳源进行生长。叠氮化物或2,4-二硝基苯酚的生长抑制水平的存在会显著抑制摄取(约90%),并且对萘的结构类似物的存在敏感。叠氮化物或2,4-二硝基苯酚的存在不会显著降低细胞内ATP水平。发现α-萘酚的存在会非竞争性抑制萘的摄取,显示出0.041微摩尔的抑制常数(Ki)。得出的结论是,荧光假单胞菌利用萘的第一步是通过特定的能量连接转运系统将其转运到细胞中。