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用于通过颜色变化对酚类化合物进行现场检测的冻干重组细菌。

Freeze-dried recombinant bacteria for on-site detection of phenolic compounds by color change.

作者信息

Shin Hae Ja, Park Hoo Hwi, Lim Woon Ki

机构信息

Environmental Engineering Major, Division of Applied Bioengineering, Dongseo University, Busan 617-716, Republic of Korea.

出版信息

J Biotechnol. 2005 Sep 22;119(1):36-43. doi: 10.1016/j.jbiotec.2005.06.002.

Abstract

We herein report the development of a recombinant bacterial biosensor for the rapid and easy detection of phenolic compounds in the field. A plasmid was designed to encode a beta-galactosidase reporter gene under the control of capR, an activator involved in phenolic compound degradation. The construct was transformed into Escherichia coli, and transformed cells were stored after being freeze-dried in the presence of sucrose. For detection of phenolic compounds, the cells were rehydrated, and used instantly, without any growth step. In the presence of 0.1 microM-10mM phenol, we observed a red color from hydrolysis of chlorophenol red beta-D-galactopyranoside (CPRG) or an indigo color from hydrolysis of X-galactopyranoside (X-gal). Other phenolic compounds could be detected by this system, including catechol, 2-methylphenol, 2-chlorophenol, 3-methylphenol, 2-nitrophenol, and 4-chlorophenol. These results suggest that this novel bacteria biosensor may be useful for easy, on-site detection of phenolic compounds without the need for unwieldy equipment or sample pretreatment. Indeed, biosensor systems involving beta-galactosidase-expressing freeze-dried recombinant bacteria could prove useful for the in situ detection of many more compounds in the future.

摘要

我们在此报告了一种重组细菌生物传感器的开发,用于在现场快速简便地检测酚类化合物。设计了一种质粒,以在capR(一种参与酚类化合物降解的激活剂)的控制下编码β-半乳糖苷酶报告基因。将构建体转化到大肠杆菌中,转化后的细胞在蔗糖存在下冻干后储存。为了检测酚类化合物,将细胞复水并立即使用,无需任何生长步骤。在存在0.1微摩尔至10毫摩尔苯酚的情况下,我们观察到氯酚红β-D-吡喃半乳糖苷(CPRG)水解产生红色,或X-吡喃半乳糖苷(X-gal)水解产生靛蓝色。该系统还可以检测其他酚类化合物,包括儿茶酚、2-甲基苯酚、2-氯苯酚、3-甲基苯酚、2-硝基苯酚和4-氯苯酚。这些结果表明,这种新型细菌生物传感器可能有助于在无需笨重设备或样品预处理的情况下,轻松地在现场检测酚类化合物。事实上,涉及表达β-半乳糖苷酶的冻干重组细菌的生物传感器系统未来可能被证明对原位检测更多化合物有用。

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