Citation: Wenchao Chen, Xiaoting Niu, Xinyu Liang, Guanghao Zhu, Xue Zhang, Qing Gong, Cen Xie, Guangbo Ge, Ping Wang, Yuqi He. Rationally engineered a universal far-red fluorogenic substrate of bile salt hydrolase for functional imaging and inhibitor screening[J]. Chinese Chemical Letters, ;2026, 37(4): 111320. doi: 10.1016/j.cclet.2025.111320 shu

Rationally engineered a universal far-red fluorogenic substrate of bile salt hydrolase for functional imaging and inhibitor screening

Wenchao Chen ^{a, b, 1} ,
Xiaoting Niu ^{b, 1} ,
Xinyu Liang ^c ,
Guanghao Zhu ^b ,
Xue Zhang ^b ,
Qing Gong ^b ,
Cen Xie ^c ,
Guangbo Ge ^{b, *,} ,
Ping Wang ^{a, b, *,} ,
Yuqi He ^{a, *,}

a.
Guizhou Engineering Research Center of Industrial Key-technology for Dendrobium Nobile, Guizhou Engineering Research Center for Orchid Medicinal Plant Breeding and Efficient Application, Zunyi Medical University, Zunyi 563000, China
b.
State Key Laboratory of Discovery and Utilization of Functional Components in Traditional Chinese Medicine, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
c.
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China

geguangbo@shutcm.edu.cn

pwang@shutcm.edu.cn

yqhe.pharm@foxmail.com

Received Date: 26 February 2025
Revised Date: 28 April 2025
Accepted Date: 14 May 2025
Available Online: 15 May 2025

Figures(5)

Bile salt hydrolase (BSH), a gatekeeper enzyme in bile acid metabolism, regulates the host's bile acid profile and is closely associated with various metabolic diseases. However, suitable methods for measuring its activity in living systems remain scarce. Herein, a novel far-red fluorogenic substrate (CA-ABEI) for BSH was designed and developed by conjugating cholic acid with an aminocoumarin fluorophore. Under physiological conditions, CA-ABEI can be rapidly hydrolyzed by BSH from various bacterial sources to form ABEI, triggering strong fluorescence enhancement at 620 nm. Specifically activated by BSH, CA-ABEI enables accurate detection of BSH activity in biospecimens, including pure enzymes, bacteria and intact fecal slurries, and the first bioimaging of BSH activity in both BSH-expressing engineered Escherichia coli and natural intestinal microbiota. Moreover, a high-throughput screening platform was established using CA-ABEI, enabling the evaluation of BSH inhibitory effects from 96 herbal extracts. Pu-erh tea emerged as a potent BSH inhibitor and its active components were subsequently characterized, aiding the discovery of novel BSH inhibitors. Collectively, CA-ABEI proved to be a powerful tool for monitoring BSH activity in complex biological systems with value for exploring physiological functions and rapid screening of inhibitors.
- Bile salt hydrolase,
- Fluorescent probe,
- Coumarin,
- High-throughput screening,
- Bioimaging
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