A real time S1 assay at neutral pH based on graphene oxide quenched fluorescence probe

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Abstract

As the extracellular nuclease of Aspergillus, S1 nuclease can split single and double-stranded DNA into oligo- or mononucleotides, while preferentially digests single-stranded nucleic acids. Furthermore, the existence of S1 can be the standard to identify Aspergillus and used to evaluate the severity of Aspergillosis. Herein, a simple and sensitive fluorescent sensing platform for S1 assay was developed based on the S1-induced DNA strand scission and the difference in affinity of graphene oxide (GO) for single-stranded DNA containing different bases. This platform was applied to monitor S1 activity and study the kinetics in real time. Results indicated that the detection limit is 0.5 U/mL. The Km and kcat at 45°C, are 1.4±0.12μM and 0.6min-1, respectively. Moreover, by monitoring the effect of chemical drugs on S1 activity, we found that 2 mM of erythromycin, sodium penicillin, carbenicillin disodium and ampicillin can inhibit S1 activity about 8%, 60%, 61% and 66%, respectively, while gentamycin sulfate is a stimulator. Overall, the assay platform based on graphene oxide quenched fluorescence probe is successfully constructed to study the enzymatic activity of S1 and used for screening antibiotics.

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APA

Xu, W., Peng, L., Li, B., Xie, Z., Tong, C., & Liu, B. (2016). A real time S1 assay at neutral pH based on graphene oxide quenched fluorescence probe. Sensing and Bio-Sensing Research, 7, 42–47. https://doi.org/10.1016/j.sbsr.2015.12.002

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