Label-Free Visualization of Carbapenemase Activity in Living Bacteria

Ye Zhang; Jin-E Lei; Yuan He; Jianhua Yang; Wenjing Wang; Abdul Wasey; Jiru Xu; Yue Lin; Haiming Fan; Guangyin Jing; Ce Zhang; Yi Jin

doi:10.1002/anie.201810834

Label-Free Visualization of Carbapenemase Activity in Living Bacteria

Ye Zhang, Jin-E Lei, Yuan He, Jianhua Yang, Wenjing Wang, Abdul Wasey, Jiru Xu, Yue Lin, Haiming Fan, Guangyin Jing, Ce Zhang, Yi Jin

Chemical Biology and Biological Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

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.

Original language	English
Pages (from-to)	17120-17124
Number of pages	5
Journal	Angewandte Chemie International Edition
Volume	57
Issue number	52
DOIs	https://doi.org/10.1002/anie.201810834
Publication status	Published - 2018

Keywords

calorimetry
bacterial detection
carbapenemase
Enterobacteriaceae
enzyme kinetics

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10.1002/anie.201810834

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title = "Label-Free Visualization of Carbapenemase Activity in Living Bacteria",

abstract = "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.",

keywords = "calorimetry, bacterial detection, carbapenemase, Enterobacteriaceae, enzyme kinetics",

author = "Ye Zhang and Jin-E Lei and Yuan He and Jianhua Yang and Wenjing Wang and Abdul Wasey and Jiru Xu and Yue Lin and Haiming Fan and Guangyin Jing and Ce Zhang and Yi Jin",

note = "https://doi.org/10.1002/anie.201810834",

year = "2018",

doi = "10.1002/anie.201810834",

language = "English",

volume = "57",

pages = "17120--17124",

journal = "Angewandte Chemie International Edition",

issn = "1433-7851",

publisher = "John Wiley & Sons Ltd",

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TY - JOUR

T1 - Label-Free Visualization of Carbapenemase Activity in Living Bacteria

AU - Zhang, Ye

AU - Lei, Jin-E

AU - He, Yuan

AU - Yang, Jianhua

AU - Wang, Wenjing

AU - Wasey, Abdul

AU - Xu, Jiru

AU - Lin, Yue

AU - Fan, Haiming

AU - Jing, Guangyin

AU - Zhang, Ce

AU - Jin, Yi

N1 - https://doi.org/10.1002/anie.201810834

PY - 2018

Y1 - 2018

N2 - 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.

AB - 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.

KW - calorimetry

KW - bacterial detection

KW - carbapenemase

KW - Enterobacteriaceae

KW - enzyme kinetics

U2 - 10.1002/anie.201810834

DO - 10.1002/anie.201810834

M3 - Article

SN - 1433-7851

VL - 57

SP - 17120

EP - 17124

JO - Angewandte Chemie International Edition

JF - Angewandte Chemie International Edition

IS - 52

ER -

Label-Free Visualization of Carbapenemase Activity in Living Bacteria

Abstract

Keywords

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