A new method for the in situ assay of α-glucosidase activity and inhibitor screening through an enzyme substrate-mediated DNA hybridization chain reaction. (31st May 2019)

• Record Type: Journal Article
• Authors: Yue, Xiquan; Su, Lihong; Chen, Xu; Liu, Junfeng; Zheng, Longpo; Zhang, Juan
• Is Part Of: New journal of chemistry. Volume 43:Number 24(2019)
• Page Start: 9458
• Page End: 9465
• Publication Date: 2019-05-31
• Subjects: Chemistry -- Periodicals; Chimie -- Périodiques; 540
• DOI: 10.1039/c9nj01868a
• Languages: English
• ISSNs: 1144-0546

Abstract

Abstract : The strategy is based on small molecule-mediated hybridization chain reaction. Abstract : In this work, a new strategy has been proposed for the in situ analysis of α-glucosidase (α-Glu) based on the small molecule-mediated DNA hybridization chain reaction (HCR). As an important digestion enzyme, α-Glu could catalyze the hydrolysis of the substrate para -aminophenyl-α-d -glucopyranoside (pAPG) into para -aminophenyl (pAP) and α-d -glucopyranoside. Among them, pAPG with cis -diols groups could coordinate with the boronic acid-tagged initiation chain to drive HCR around the assembled magnetic nanospheres (AMNSs). Nevertheless, the product of the catalytic reaction pAP did not have the capability to capture the boronic acid-tagged initiation chain onto the surface of AMNSs, due to which HCR did not occur. Thus, the α-Glu activity was correlated with the extent of HCR around AMNSs and it could be tested in situ in a cell medium with the aid of a magnet. The enzyme activity was analyzed over a wide linear range from 0.4 U mL −1 to 1.7 U mL −1 with the lowest detection limit of 0.023 U mL −1 . Furthermore, the established method could also be successfully utilized for the in situ evaluation of enzyme inhibitors including gallic acid and quercetin. This study may provide a new insight for the development of a new method for the in situ analysis of α-Glu in a cell medium.