Please use this identifier to cite or link to this item: http://hdl.handle.net/10397/81714
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Title: Thermostable beta-lactamase mutant with its active site conjugated with fluorescein for efficient beta-lactam antibiotic detection
Authors: Au, HW 
Tsang, MW 
So, PK 
Wong, KY 
Leung, YC 
Issue Date: 2019
Source: ACS omega, 27 Nov. 2019, v. 4, no. 24, p. 20493-20502
Abstract: Monitoring the beta-lactam antibiotic level has been an important task in food industry and clinical practice. Here, we report the development of a fluorescent PenP beta-lactamase, PenP-E166Cf/N170Q for efficient beta-lactam antibiotic detection. It was constructed by covalently attaching fluorescein onto the active-site entrance of a thermostable E166Cf/N170Q mutant of a Bacillus licheniformis PenP beta-lactamase. It gave a fluorescence turn-on signal toward various beta-lactam antibiotics, where the fluorescence enhancement was attributed to the acyl-enzyme complex formed between PenP-E166Cf/N170Q and the beta-lactam antibiotic. It demonstrated enhanced signal stability over its parental PenP-E166Cf because of the suppressed hydrolytic activity by the N170Q mutation. Compared with our previously constructed PenPC-E166Cf biosensor, PenP-E166Cf/N170Q was more thermostable and advanced in detecting beta-lactams in terms of response time, signal stability, and detection limit. Positive fluorescence signals generated by E166Cf/N170Q in response to the penicillin-containing milk and mouse serum illustrated the feasibility of the biosensor for antibiotic detection in real samples. Taken together, our findings suggest the potential application of PenP-E166Cf/N170Q in biosensing beta-lactam antibiotics.
Publisher: American Chemical Society
Journal: ACS omega 
ISSN: 2470-1343
DOI: 10.1021/acsomega.9b02211
Rights: Copyright © 2019 American Chemical Society
This is an open access article published under an ACS AuthorChoice License (https://pubs.acs.org/page/policy/authorchoice_termsofuse.html), which permits copying and redistribution of the article or any adaptations for non-commercial purpose
The following publication Au, H. W., Tsang, M. W., So, P. K., Wong, K. Y., & Leung, Y. C. (2019). Thermostable beta-lactamase mutant with its active site conjugated with fluorescein for efficient beta-lactam antibiotic detection. ACS Omega, 4(24), 20493-20502 is available at https://dx.doi.org/10.1021/acsomega.9b02211
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