Luque-Almagro Victor M, Huertas María J, Roldán M Dolores, Moreno-Vivián Comrado, Martínez-Luque Manuel, Blasco Rafael, Castillo Francisco
Departamento de Bioquímica y Biología Molecular, Campus de Rabanales, Edificio Severo Ochoa, Universidad de Córdoba, 14071 Córdoba, Spain.
Environ Microbiol. 2007 Jun;9(6):1541-9. doi: 10.1111/j.1462-2920.2007.01274.x.
Two-dimensional (2-D) electrophoresis approach has been used to test protein expression changes in response to cyanide in the alkaliphilic bacterium Pseudomonas pseudoalcaligenes CECT5344. This is a cyanide-assimilating strain which also grows in media containing cyanide-enriched effluent from the jewellery industry. The bacterium efficiently uses this residue as the sole nitrogen source for aerobic growth under alkaline pH with negligible nitrogen losses as HCN. Cell-free extracts isolated from P. pseudoalcaligenes grown with a jewellery residue, free cyanide or ammonium chloride as nitrogen source were subjected to 2-D electrophoresis and the spot patterns were examined to determine differential protein expression. Electrophoretic plates exhibiting an average of 1000 spots showed significant differences in the expression of about 44 proteins depending on the nitrogen source. Some of these protein spots were analysed by Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). Characterization of five of these proteins reveals that cyanide shock induces proteins related to iron acquisition, regulation of nitrogen assimilation pathways and oxidative stress repairing and protection.
二维(2-D)电泳方法已被用于检测嗜碱细菌假产碱假单胞菌CECT5344中蛋白质表达对氰化物的响应变化。这是一种能同化氰化物的菌株,它也能在含有珠宝行业富含氰化物废水的培养基中生长。该细菌能有效地利用这种残留物作为碱性pH条件下有氧生长的唯一氮源,以HCN形式损失的氮可忽略不计。从以珠宝残留物、游离氰化物或氯化铵作为氮源生长的假产碱假单胞菌中分离出的无细胞提取物进行二维电泳,并检查斑点模式以确定差异蛋白质表达。平均显示1000个斑点的电泳板表明,根据氮源不同,约44种蛋白质的表达存在显著差异。其中一些蛋白质斑点通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)进行了分析。对其中五种蛋白质的表征表明,氰化物冲击会诱导与铁获取、氮同化途径调节以及氧化应激修复和保护相关的蛋白质。