Stahl Antje, Lazar Alexandra I, Muchemu Veronica N, Nau Werner M, Ullrich Matthias S, Hennig Andreas
Department of Life Sciences and Chemistry, Jacobs University Bremen, Campus Ring 1, 28759, Bremen, Germany.
Anal Bioanal Chem. 2017 Nov;409(27):6485-6494. doi: 10.1007/s00216-017-0593-2. Epub 2017 Sep 6.
Steroids have been identified as endocrine-disrupting agents, which are thought to impact the fertility of aquatic organisms and may even have direct effects on humans. The removal of steroids from wastewater is therefore essential, and this is most efficiently achieved by microbial treatment. We report herein a simple fluorescent method to identify microorganisms that are capable of steroid degradation and to optimize the conditions for steroid removal. The method is based on the supramolecular macrocycle cucurbit[8]uril (CB8), which can bind either the fluorescent dye berberine or a steroid in their inner cavity. In absence of steroid, the cavity is free to bind the dye, leading to a strong increase in fluorescence. In contrast, in the presence of steroid, the dye is displaced into the bulk solution. This principle affords a stable (no thermal or photodegradation was noted), fluorescent chemosensor (excitation ca. 450 nm, maximum emission at 525 nm), which can detect testosterone at concentrations > 0.7 μM. We show that this displacement principle can be applied to follow the removal of micromolar concentrations of the steroid testosterone from a bacterial culture of Buttiauxella sp. S19-1. The reliability of the chemosensor in screening applications is demonstrated by an excellent Z-factor, which was in the range of 0.52 to 0.74 for all experiments carried out with this assay. Graphical abstract Steroid depletion by bacterial cultures can be followed by fluorescence spectroscopy using a supramolecular chemosensor based on berberine and cucurbit[8]uril.
类固醇已被确认为内分泌干扰物，被认为会影响水生生物的繁殖力，甚至可能对人类产生直接影响。因此，从废水中去除类固醇至关重要，而通过微生物处理能最有效地实现这一点。我们在此报告一种简单的荧光方法，用于识别能够降解类固醇的微生物并优化类固醇去除条件。该方法基于超分子大环葫芦[8]脲（CB8），它可以在其内腔中结合荧光染料黄连素或类固醇。在没有类固醇的情况下，内腔可自由结合染料，导致荧光强烈增强。相反，在有类固醇的情况下，染料会被置换到本体溶液中。这一原理提供了一种稳定的（未观察到热降解或光降解）荧光化学传感器（激发波长约450nm，最大发射波长在525nm），它能够检测浓度大于0.7μM的睾酮。我们表明，这种置换原理可用于跟踪从布氏菌属菌株S19-1的细菌培养物中去除微摩尔浓度的类固醇睾酮的过程。通过出色的Z因子证明了该化学传感器在筛选应用中的可靠性，在使用该测定法进行的所有实验中，Z因子范围为0.52至0.74。图形摘要 细菌培养物中类固醇的消耗可以通过使用基于黄连素和葫芦[8]脲的超分子化学传感器的荧光光谱法进行跟踪。
