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Label‐Free Visualization of Carbapenemase Activity in Living Bacteria
Author(s) -
Zhang Ye,
Lei JinE,
He Yuan,
Yang Jianhua,
Wang Wenjing,
Wasey Abdul,
Xu Jiru,
Lin Yue,
Fan Haiming,
Jing Guangyin,
Zhang Ce,
Jin Yi
Publication year - 2018
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201810834
Subject(s) - bacteria , imipenem , substrate (aquarium) , chemistry , enterobacteriaceae , kinetics , enzyme , computational biology , biochemistry , microbiology and biotechnology , combinatorial chemistry , biology , pseudomonas aeruginosa , escherichia coli , genetics , ecology , physics , quantum mechanics , gene
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.