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High-throughput fluorescence screening assay for the identification and comparison of antimicrobial peptides’ activity on various yeast species

SYSNO ASEP	0461653
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	High-throughput fluorescence screening assay for the identification and comparison of antimicrobial peptides’ activity on various yeast species
Author(s)	Kodedová, Marie (FGU-C)_{RID, ORCID} Sychrová, Hana (FGU-C)_{RID, ORCID}
Source Title	Journal of Biotechnology. - : Elsevier - ISSN 0168-1656 Roč. 233, Sep 10 (2016), s. 26-33
Number of pages	8 s.
Language	eng - English
Country	NL - Netherlands
Keywords	diS-C3(3) assay ; antimicrobial peptide ; Candida ; membrane potential ; microplate reader
Subject RIV	EE - Microbiology, Virology
R&D Projects	TA04010638 GA TA ČR - Technology Agency of the Czech Republic (TA ČR)
	ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
Institutional support	FGU-C - RVO:67985823
UT WOS	000380819300004
EID SCOPUS	84978253165
DOI	10.1016/j.jbiotec.2016.06.023
Annotation	New antifungal compounds that circumvent the resistance of the pathogen by directly damaging yeast cell surface structures are promising agents for the treatment of fungal infections, due to their different mechanism of action from current clinically used antifungal drugs. We present here a rapid and cost-effective fluorescence method suitable for identifying new potent drugs that directly target yeast cell surface structures, causing cell permeabilization and thus bypassing the multidrug resistance mechanisms of pathogens. The fluorescence assay enabled us to detect with high sensitivity damage to the Candida plasma membrane (its hyperpolarization and permeabilization) as a result of short-term exposure to the antifungal compounds. Results can be obtained in 1–2 h with minimal effort and consumption of the tested compounds, also 96 samples can be analysed simultaneously. We used this method to study antimicrobial peptides isolated from the venom of bees and their synthetic analogs, compare the potency of the peptides and determine their minimal effective concentrations. The antimicrobial peptides were able to kill yeast cells at low concentrations within a 15-min treatment, the LL-III peptide exhibited a broad spectrum of antifungal activity on various Saccharomyces, pathogenic Candida and osmotolerant yeast species.
Workplace	Institute of Physiology
Contact	Lucie Trajhanová, lucie.trajhanova@fgu.cas.cz, Tel.: 241 062 400
Year of Publishing	2017

Number of the records: 1