Yoon Hey Young, Kim Hyung Jun, Jang Soojin, Hong Jong-In
Department of Chemistry, College of Natural Sciences, Seoul National University, Seoul, 08826, Republic of Korea.
Antibacterial Resistance Research Laboratory, Department of Discovery Biology, Institute Pasteur Korea, 16 Daewangpangyo-ro 712 beon-gi, Bundang-gu, Seongnam, Gyeonggi-do, 13488, Republic of Korea.
AMB Express. 2017 Dec;7(1):150. doi: 10.1186/s13568-017-0449-3. Epub 2017 Jul 11.
Bacterial arylsulfatases are crucial to biosynthesis in many microorganisms, as bacteria often utilize aryl sulfates as a source of sulfur. The bacterial sulfatases are associated with pathogenesis and are applied in many areas such as industry and agriculture. We developed an activity-based probe 1 for detection of bacterial sulfatase activity through liquid- and solid-phase colony-based assays. Probe 1 is hydrolyzed by sulfatase to generate fluorescent N-methyl isoindole, which is polymerized to form colored precipitates. These fluorescent and colorimetric properties of probe 1 induced upon treatment of sulfatases were successfully utilized for liquid-phase sulfatase activity assays for colonies and lysates of Klebsiella aerogenes, Mycobacterium avium and Mycobacterium smegmatis. In addition, probe 1 allowed solid-phase colony-based assays of K. aerogenes through the formation of insoluble colored precipitates, thus enabling accurate staining of target colonies under heterogeneous conditions.
细菌芳基硫酸酯酶对许多微生物的生物合成至关重要，因为细菌常常利用芳基硫酸盐作为硫源。细菌硫酸酯酶与发病机制相关，并应用于工业和农业等许多领域。我们开发了一种基于活性的探针1，通过基于液相和固相菌落的检测方法来检测细菌硫酸酯酶活性。探针1被硫酸酯酶水解生成荧光N-甲基异吲哚，后者聚合形成有色沉淀。探针1经硫酸酯酶处理后产生的这些荧光和比色特性被成功用于产气克雷伯菌、鸟分枝杆菌和耻垢分枝杆菌菌落及裂解物的液相硫酸酯酶活性检测。此外，探针1通过形成不溶性有色沉淀实现了产气克雷伯菌基于固相菌落的检测，从而能够在异质条件下对目标菌落进行准确染色。
