La Trobe

A novel red-emitting aggregation-induced emission probe for determination of β-glucosidase activity

journal contribution
posted on 2025-02-06, 03:07 authored by Bicheng YaoBicheng Yao, Jiamin ZhaoJiamin Zhao, Siyang DingSiyang Ding, Marie-Claire Giel, G Zhang, D Ding, Y Tang, Zhe Weng, Yuning HongYuning Hong
β-Glucosidase (β-Glu) is a ubiquitous enzyme which has multiple roles in medical diagnosis, food production, agriculture, etc. Existing β-Glu assays have limitations such as complex operation, long running time, and high background noise. Here we report a red-emissive probe TBPG for measuring the activity of β-Glu. The probe was synthesized through conjugating a β-Glu targeting glucoside to an aggregation-induced emission (AIE) fluorophore. In the presence of β-Glu, TBPG was hydrolyzed and exhibited a fluorescence turn-on process. The detection conditions including time, temperature, pH value, buffer, and probe concentration were optimized systematically. Afterwards, fluorescence titration was conducted showing an excellent linearity (R2 = 0.998), a wide linear dynamic range (0–5.0 U/mL), and a limit of detection as low as 0.6 U/L. The detection specificity and ion interference were evaluated by adding various biological species and ions to probe without or with β-Glu. Next, we demonstrate the applicability of probe TBPG in determining the β-Glu activity in living cells using confocal microscopy and flow cytometry. Finally, this newly established assay was applied to real soil samples. Comparable results were obtained as the commercial assay, manifesting its great potential in soil enzyme analysis.

Funding

This work was supported by grants to Y.H. (Australian Research Council FT210100271 and Australia-China Science and Research Fund-Joint Research Centre on Personal Health Technologies ACSRF65777) and to Z.H.W. (La Trobe University's Research Focus Area in Securing Food, Water and the Environment, SFWE RFA 2000004295 & SFWE RFA 2000004349) .

History

Publication Date

2023-04-01

Journal

Biomaterials

Volume

295

Article Number

122046

Pagination

11p.

Publisher

Elsevier

ISSN

0142-9612

Rights Statement

© 2023 Elsevier Ltd. All rights reserved. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/

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