Label-free visualization of carbapenemase activity in living bacteria

Zhang, Ye, Lei, Jin-E, He, Yuan, Yang, Jianhua, Wang, Wenjing, Wasey, Abdul, Xu, Jiru, Lin, Yue, Fan, Haiming, Jing, Guangyin, Zhang, Ce and Jin, Yi ORCID: https://orcid.org/0000-0002-6927-4371 2018. Label-free visualization of carbapenemase activity in living bacteria. Angewandte Chemie International Edition 57 (52) , pp. 17120-17124. 10.1002/anie.201810834

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Abstract

Evaluating enzyme activity intracellularly on natural substrates is a significant experimental challenge in biomedical research. We report a label‐free method for real‐time monitoring of the catalytic behavior of class A, B, and D carbapenemases in live bacteria based on measurement of heat changes. By this means, novel biphasic kinetics for class D OXA‐48 with imipenem as substrate is revealed, providing a new approach to detect OXA‐48‐like producers. This in‐cell calorimetry approach offers major advantages in the rapid screening (10 min) of carbapenemase‐producing Enterobacteriaceae from 142 clinical bacterial isolates, with superior sensitivity (97 %) and excellent specificity (100 %) compared to conventional methods. As a general, label‐free method for the study of living cells, this protocol has potential for application to a wider range and variety of cellular components and physiological processes.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Chemistry
Publisher:	Wiley
ISSN:	1433-7851
Date of First Compliant Deposit:	10 December 2018
Date of Acceptance:	31 October 2018
Last Modified:	12 Nov 2024 18:45
URI:	https://orca.cardiff.ac.uk/id/eprint/117523

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