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- Title
Label‐Free Visualization of Carbapenemase Activity in Living Bacteria.
- Authors
Zhang, Ye; Lei, Jin‐E; He, Yuan; Yang, Jianhua; Wang, Wenjing; Wasey, Abdul; Xu, Jiru; Lin, Yue; Fan, Haiming; Jing, Guangyin; Zhang, Ce; Jin, Yi
- 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.
- Subjects
CARBAPENEMASE; ENZYME activation; BIOLOGICAL research; CHEMICAL kinetics; CALORIMETRY
- Publication
Angewandte Chemie, 2018, Vol 130, Issue 52, p17366
- ISSN
0044-8249
- Publication type
Article
- DOI
10.1002/ange.201810834