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http://hdl.handle.net/10397/89163
Title: | BADAN-conjugated ?-lactamases as biosensors for ?-lactam antibiotic detection | Authors: | Au, HW Tsang, MW Chen, YW So, PK Wong, KY Leung, YC |
Issue Date: | Oct-2020 | Source: | PLoS one, 30 Oct. 2020, v. 15, e0241594, p. 1-13 | Abstract: | β-Lactam antibiotic detection has significant implications in food safety control, environmental monitoring and pharmacokinetics study. Here, we report the development of two BADAN-conjugated β-lactamases, E166Cb and E166Cb/N170Q, as sensitive biosensors for β-lactam antibiotic detection. These biosensors were constructed by coupling an environment-sensitive BADAN probe onto location 166 at the active site of the PenP β-lactamase E166C and E166C/N170Q mutants. They gave fluorescence turn-on signals in response to β-lactam antibiotics. Molecular dynamics simulation of E166Cb suggested that the turn-on signal might be attributed to a polarity change of the microenvironment of BADAN and the removal of the fluorescence quenching effect on BADAN exerted by a nearby Tyr-105 upon the antibiotic binding. In the detection of four β-lactams (penicillin G, penicillin V, cefotaxime and moxalactam), both E166Cb and E166Cb/N170Q delivered signal outputs in an antibiotic-concentration dependent manner with a dynamic range spanning from 10 nM to 1 μM. Compared to E166Cb, E166Cb/N170Q generally exhibited more stable signals owing to its higher deficiency in hydrolyzing the antibiotic analyte. The overall biosensor performance of E166Cb and E166Cb/N170Q was comparable to that of their respective fluorescein-modified counterparts, E166Cf and E166Cf/N170Q. But comparatively, the BADAN-conjugated enzymes showed a higher sensitivity, displayed a faster response in detecting moxalactam and a more stable fluorescence signals towards penicillin G. This study illustrates the potential of BADAN-conjugated β-lactamases as biosensing devices for β-lactam antibiotics. | Publisher: | Public Library of Science | Journal: | PLoS one | EISSN: | 1932-6203 | DOI: | 10.1371/journal.pone.0241594 | Rights: | © 2020 Au et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The following publication Au H-W, Tsang M-W, Chen YW, So P-K, Wong K-Y, Leung Y-C (2020) BADAN-conjugated β-lactamases as biosensors for β-lactam antibiotic detection. PLoS ONE 15(10): e0241594 is available at https://dx.doi.org/10.1371/journal.pone.0241594 |
Appears in Collections: | Journal/Magazine Article |
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