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In Vitro Gene Expression-Coupled Bacterial Cell Chip for Screening Species-Specific Antimicrobial Enzymes

Authors
Kwon, Seok-JoonKim, DomyoungLee, InseonKim, JungbaeDordick, Jonathan S.
Issue Date
8월-2017
Publisher
WILEY
Keywords
lysins; bacterial cell chip; high-throughput screening; in vitro TNT; SA1
Citation
BIOTECHNOLOGY AND BIOENGINEERING, v.114, no.8, pp.1648 - 1657
Indexed
SCIE
SCOPUS
Journal Title
BIOTECHNOLOGY AND BIOENGINEERING
Volume
114
Number
8
Start Page
1648
End Page
1657
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/82772
DOI
10.1002/bit.26300
ISSN
0006-3592
Abstract
Targeting infectious bacterial pathogens is important for reducing the evolution of antibiotic-resistant bacteria and preserving the endogenous human microbiome. Cell lytic enzymes including bacteriophage endolysins, bacterial autolysins, and other bacteriolysins are useful antibiotic alternatives due to their exceptional target selectivity, which may be used to lysins rapidly kill target bacteria and their high specificity permit the normal commensal microflora to be left undisturbed. Genetic information of numerous lysins is currently available, but the identification of their antimicrobial function and specificity has been limited because most lysins are often poorly expressed and exhibit low solubilities. Here, we report the development of bacterial cell chip for rapidly accessing the function of diverse genes that are suggestive of encoding lysins. This approach can be used to evaluate rapidly the species-specific antimicrobial activity of diverse lysins synthesized from in vitro transcription and translation (TNT) of plasmid DNA. In addition, new potent lysins can be assessed that are not expressed in hosts and display low solubility. As a result of evaluating the species-specific antimicrobial function of 11 (un)known lysins with an in vitro TNT-coupled bacterial cell chip, a potent recombinant lysin against Staphylococcus strains, SA1, was identified. The SA1 was highly potent against not only S. aureus, but also both lysostaphin-resistant S. simulans and S. epidermidis cells. To this end, the SA1 may be applicable to treat both methicillin-resistant S. aureus (MRSA) and lysostaphin-resistant MRSA mutants. (c) 2017 Wiley Periodicals, Inc.
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공과대학 (화공생명공학과)
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