title: In vitro gene expression-coupled bacterial cell chip for screening species-specific antimicrobial enzymes.
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. Biotechnol. Bioeng. 2017;114: 1648-1657. © 2017 Wiley Periodicals, Inc.
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, New York, 12180.
authors.forename: Seok-Joon
authors.initials: SJ
authors.lastname: Kwon
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, New York, 12180.
authors.forename: Domyoung
authors.initials: D
authors.lastname: Kim
authors.affiliation: Department of Chemical and Biological Engineering, Korea University, Seoul, 02841, Republic of Korea.
authors.forename: Inseon
authors.initials: I
authors.lastname: Lee
authors.affiliation: Department of Chemical and Biological Engineering, Korea University, Seoul, 02841, Republic of Korea.
authors.forename: Jungbae
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, New York, 12180.
authors.forename: Jonathan S
authors.identifier: 0000-0001-7802-3702
authors.initials: JS
authors.lastname: Dordick
mesh_terms: D018407:Bacterial Physiological Phenomena / Q000187:drug effects*; D001426:Bacterial Proteins / Q000008:administration & dosage*; D001681:Biological Assay / Q000295:instrumentation*; D002470:Cell Survival / Q000187:drug effects; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation*; D004798:Enzymes / Q000008:administration & dosage*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D020869:Gene Expression Profiling / Q000295:instrumentation*; D018511:Systems Integration; D046888:Tissue Array Analysis / Q000295:instrumentation
chemical_list: D001426:Bacterial Proteins; D004798:Enzymes
keywords: SA1; bacterial cell chip; high-throughput screening; in vitro TNT; lysins