Sar P, Kazy S K, Asthana R K, Singh S P
Algal Research Laboratory, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005, India.
Curr Microbiol. 1998 Nov;37(5):306-11. doi: 10.1007/s002849900383.
Pseudomonas aeruginosa cells growing in minimal medium were 40-fold more sensitive to Ni2+ than cells growing in enriched medium, suggesting a possible protective role of medium ingredients. Likewise, cells pre-grown in enriched medium showed a high Km (6.15 mM) and increased Ni2+ uptake (950 nmol mg-1 protein, 1h) over cells pre-sown in minimal medium (Km, 0.48 mM; 146 nmol mg-1 protein, 1 h). The overall pattern indicates that cells pre-grown in enriched medium were characterized by having lowered affinity towards Ni2+ than those with minimal medium background. The enhanced Ni2+ uptake by enriched medium-grown cells can be correlated with the improved metabolic state of the cells. Ni2+ uptake was optimum at neutrality (pH 7.0). A major Ni2+ transport system was competitively inhibited by Mg2+, Zn2+, Cd2+, or Co2+ (400 microM each). Noticeably, a minor Ni2+ transport pathway was still operative even in the higher concentration range of Mg2+ (4 mM and 40 mM). The stimulation of Ni2+ uptake monitored in the presence of different carbon sources (0.5% wt/vol, each) showed the sequence: glucose (1.6-fold) > phenol = gallic acid (1.5-fold). Succinate, in comparison, reduced Ni2+ uptake (0.5-fold) possibly because of its acting as a metal chelator as well. Sensitivity of Ni2+ transport towards methyl viologen, azide, 2-4 DNP, and DCCD suggested that transport was energy-linked.
在基本培养基中生长的铜绿假单胞菌细胞对Ni2+的敏感性比在富集培养基中生长的细胞高40倍,这表明培养基成分可能具有保护作用。同样,在富集培养基中预培养的细胞显示出较高的Km值(6.15 mM),并且与在基本培养基中预培养的细胞(Km,0.48 mM;146 nmol mg-1蛋白质,1小时)相比,Ni2+摄取量增加(950 nmol mg-1蛋白质,1小时)。总体模式表明,在富集培养基中预培养的细胞对Ni2+的亲和力低于具有基本培养基背景的细胞。富集培养基中生长的细胞对Ni2+摄取的增强可能与细胞代谢状态的改善有关。Ni2+摄取在中性(pH 7.0)时最佳。主要的Ni2+转运系统受到Mg2+、Zn2+、Cd2+或Co2+(各400 microM)的竞争性抑制。值得注意的是,即使在较高浓度的Mg2+(4 mM和40 mM)下,次要的Ni2+转运途径仍然有效。在存在不同碳源(各0.5% wt/vol)的情况下监测到的Ni2+摄取刺激显示出以下顺序:葡萄糖(1.6倍)>苯酚 = 没食子酸(1.5倍)。相比之下,琥珀酸降低了Ni2+摄取(0.5倍),可能是因为它也作为金属螯合剂起作用。Ni2+转运对甲基紫精、叠氮化物、2,4-二硝基苯酚和二环己基碳二亚胺的敏感性表明转运与能量相关。