Inoue H, Takimura O, Fuse H, Murakami K, Kamimura K, Yamaoka Y
Marine Biological Technology Section, Chugoku National Industrial Research Institute, Hiroshima, Japan.
Appl Environ Microbiol. 2000 Aug;66(8):3492-8. doi: 10.1128/AEM.66.8.3492-3498.2000.
Triphenyltin (TPT)-degrading bacteria were screened by a simple technique using a post-column high-performance liquid chromatography using 3,3',4',7-tetrahydroxyflavone as a post-column reagent for determination of TPT and its metabolite, diphenyltin (DPT). An isolated strain, strain CNR15, was identified as Pseudomonas chlororaphis on the basis of its morphological and biochemical features. The incubation of strain CNR15 in a medium containing glycerol, succinate, and 130 microM TPT resulted in the rapid degradation of TPT and the accumulation of approximately 40 microM DPT as the only metabolite after 48 h. The culture supernatants of strain CNR15, grown with or without TPT, exhibited a TPT degradation activity, whereas the resting cells were not capable of degrading TPT. TPT was stoichiometrically degraded to DPT by the solid-phase extract of the culture supernatant, and benzene was detected as another degradation product. We found that the TPT degradation was catalyzed by low-molecular-mass substances (approximately 1,000 Da) in the extract, termed the TPT-degrading factor. The other fluorescent pseudomonads, P. chlororaphis ATCC 9446, Pseudomonas fluorescens ATCC 13525, and Pseudomonas aeruginosa ATCC 15692, also showed TPT degradation activity similar to strain CNR15 in the solid-phase extracts of their culture supernatants. These results suggest that the extracellular low-molecular-mass substance that is universally produced by the fluorescent pseudomonad could function as a potent catalyst to cometabolite TPT in the environment.
采用一种简单技术筛选三苯基锡(TPT)降解菌,该技术使用柱后高效液相色谱法,以3,3',4',7 - 四羟基黄酮作为柱后试剂来测定TPT及其代谢产物二苯基锡(DPT)。根据其形态和生化特征,分离出的菌株CNR15被鉴定为嗜麦芽窄食单胞菌。将菌株CNR15在含有甘油、琥珀酸盐和130微摩尔TPT的培养基中培养,48小时后TPT迅速降解,仅积累了约40微摩尔的DPT作为唯一代谢产物。菌株CNR15在有或无TPT的情况下生长的培养上清液均表现出TPT降解活性,而静息细胞则不能降解TPT。培养上清液的固相提取物将TPT化学计量地降解为DPT,并检测到苯是另一种降解产物。我们发现提取物中低分子量物质(约1000道尔顿)催化TPT降解,该物质被称为TPT降解因子。其他荧光假单胞菌,嗜麦芽窄食单胞菌ATCC 9446、荧光假单胞菌ATCC 13525和铜绿假单胞菌ATCC 15692,在其培养上清液的固相提取物中也表现出与菌株CNR15相似的TPT降解活性。这些结果表明,荧光假单胞菌普遍产生的细胞外低分子量物质可能作为一种有效的催化剂在环境中对TPT进行共代谢。