Detection of protease and engineered phage-infected bacteria using peptide-graphene oxide nanosensors
Detection of protease and engineered phage-infected bacteria using peptide-graphene oxide nanosensors
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DOI：
10.1007/s00216-019-01766-6
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发表时间：
2019-05-01
影响因子：
4.3
通讯作者：
Nugen, Sam R.
中科院分区：
文献类型：
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作者：
Chen, Juhong;Nugen, Sam R.
一种肽-氧化石墨烯纳米传感器已被开发用于检测烟草蚀纹病毒（TEV）蛋白酶和感染工程噬菌体的细菌。在检测策略中，用荧光团标记肽（序列：RKRFRENLYSSCP），并使用氧化石墨烯（GO）吸附肽，同时淬灭其荧光。在TEV蛋白酶存在下，荧光肽在谷氨酰胺（Q）和丝氨酸（S）之间裂解，导致荧光信号的恢复。根据荧光强度，TEV蛋白酶的检出限为51 ng/L。此外，我们已经利用传感系统来检测细菌细胞。噬菌体，其被工程化以携带TEV蛋白酶基因，被用于感染靶细菌（大肠杆菌），导致蛋白酶的翻译和释放。这允许在10（4）CFU/mL的浓度下估计细菌。这种策略有可能被开发为多个细菌物种的多重检测平台。
A peptide-graphene oxide nanosensor has been developed to detect tobacco etch virus (TEV) protease and bacteria infected with an engineered bacteriophage. In the detection strategy, a peptide (sequence: RKRFRENLYFQSCP) is tagged with fluorophores and graphene oxide (GO) is used to adsorb the peptides while quenching their fluorescence. In the presence of TEV protease, fluoropeptides are cleaved between glutamine (Q) and serine (S), resulting in the recovery of fluorescence signal. Based on the fluorescent intensity, the detection limit of TEV protease is 51ng/L. Additionally, we have utilized the sensing system to detect bacteria cells. Bacteriophages, which were engineered to carry TEV protease genes, were used to infect target bacteria (Escherichia coli) resulting in the translation and release of the protease. This allowed the estimation of bacteria at the concentration of 10(4)CFU/mL. This strategy has the potential to be developed as a multiplex detection platform of multiple bacterial species.