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优化全细胞细菌生物传感器在静电纺丝醋酸纤维素 (CA) 和聚己内酯 (PCL) 纤维上的固定化用于砷检测。

Optimization of whole-cell bacterial bioreporter immobilization on electrospun cellulose acetate (CA) and polycaprolactone (PCL) fibers for arsenic detection.

机构信息

Department of Molecular Biology and Genetics, Middle East Technical University, 06800, Ankara, Turkey.

Department of Agricultural Biotechnology, Aydın Adnan Menderes University, 09970, Aydın, Turkey.

出版信息

Environ Monit Assess. 2023 May 13;195(6):666. doi: 10.1007/s10661-023-11227-4.

Abstract

Arsenic contamination is a critical global problem, and its widespread environmental detection is becoming a prominent issue. Herein, electrospun fibers of cellulose acetate (CA) and polycaprolactone (PCL) were successfully fabricated and used as the support material for immobilization of arsenic-sensing bacterial bioreporter for the first time. To date, no attempt has been made to immobilize fluorescent whole-cell bioreporter cells on electrospun fibers for arsenic detection. CA and PCL electrospun fibers were fabricated via traditional electrospinning technique and characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and contact angle meter. Following immobilization of the bacterial bioreporter cells, the immobilized bacteria were also characterized by viability assay using AlamarBlue. The effects of growth phase and cell concentration on the fluorescence response of fiber-immobilized arsenic bioreporters to arsenic were also investigated. After immobilization of arsenic bioreporters on 10 wt% PCL fiber, 91% of bacterial cells remained viable, while this value was 55.4% for cells immobilized on 12.5 wt% CA fiber. Bioreporter cells in the exponential growth phase were shown to be more sensitive to arsenic compared to aged cells. While both the electropsun PCL- and CA-immobilized bioreporters successfully detected 50 and 100 µg/L of arsenite (As (III)) concentrations, the PCL-immobilized bioreporter showed better fluorescence performance which should be investigated in future studies. This study helps to fill some gaps in the literature and demonstrates the potential for using electrospun fiber-immobilized arsenic whole-cell bioreporter for arsenic detection in water.

摘要

砷污染是一个全球性的重大问题,其广泛的环境检测正成为一个突出问题。在此,我们首次成功制备了醋酸纤维素(CA)和聚己内酯(PCL)的静电纺纤维,并将其用作固定砷感应细菌生物报告器的支撑材料。迄今为止,尚无尝试将荧光全细胞生物报告器细胞固定在静电纺纤维上来进行砷检测。CA 和 PCL 静电纺纤维通过传统的静电纺丝技术制备,并通过扫描电子显微镜(SEM)、傅里叶变换红外光谱(FTIR)和接触角仪进行了表征。在固定细菌生物报告器细胞后,还通过使用 AlamarBlue 进行活菌测定来对固定化细菌进行了表征。还研究了生长阶段和细胞浓度对纤维固定砷生物报告器对砷的荧光响应的影响。在 10 wt% PCL 纤维上固定砷生物报告器后,有 91%的细菌细胞保持存活,而在 12.5 wt% CA 纤维上固定时,这一数值为 55.4%。与老化细胞相比,处于指数生长期的生物报告器细胞对砷的敏感性更高。虽然静电纺 PCL 和 CA 固定的生物报告器都成功地检测到 50 和 100μg/L 的亚砷酸盐(As(III))浓度,但 PCL 固定的生物报告器表现出更好的荧光性能,这应该在未来的研究中进行探讨。本研究有助于填补文献中的一些空白,并展示了在水中使用静电纺纤维固定砷全细胞生物报告器进行砷检测的潜力。

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