Faculty of Agriculture, Utsunomiya University, 350 Minemachi, Utsunomiya 321-8505, Japan.
J Microbiol Biotechnol. 2011 Dec;21(12):1306-11. doi: 10.4014/jmb.1105.05017.
Recombinant Rhodopseudomonas palustris, harboring the carotenoid-metabolizing gene crtI (CrtIBS), and whose color changes from greenish yellow to red in response to inorganic As(III), was cultured in transparent microplate wells illuminated with a light emitting diode (LED) array. The cells were seen to grow better under near-infrared light, when compared with cells illuminated with blue or green LEDs. The absorbance ratio of 525 to 425 nm after cultivation for 24 h, which reflects red carotenoid accumulation, increased with an increase in As(III) concentrations. The detection limit of cultures illuminated with near-infrared LED was 5 microgram/l, which was equivalent to that of cultures in test tubes illuminated with an incandescent lamp. A near-infrared LED array, in combination with a microplate, enabled the simultaneous handling of multiple cultures, including CrtIBS and a control strain, for normalization by the illumination of those with equal photon flux densities. Thus, the introduction of a near-infrared LED array to the assay is advantageous for the monitoring of arsenic in natural water samples that may contain a number of unknown factors and, therefore, need normalization of the reporter event.
含有类胡萝卜素代谢基因 crtI(CrtIBS)的重组沼泽红假单胞菌,其颜色会根据无机砷(III)的存在从绿黄色变为红色,在发光二极管(LED)阵列照射的透明微孔板井中进行培养。与用蓝色或绿色 LED 照射的细胞相比,近红外光下的细胞生长得更好。培养 24 小时后,525nm 与 425nm 的吸光度比值反映了类胡萝卜素的积累量,随着 As(III)浓度的增加而增加。用近红外 LED 照射的培养物的检测限为 5 微克/升,与用白炽灯照射的试管培养物的检测限相当。近红外 LED 阵列与微孔板相结合,可以同时处理多个培养物,包括 CrtIBS 和对照菌株,通过用相同的光子通量密度对它们进行照射来实现归一化。因此,在测定中引入近红外 LED 阵列有利于监测可能含有多种未知因素的天然水样中的砷,因此需要对报告事件进行归一化。