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TY - JOUR

T1 - Isolation of persister cells of bacillus subtilis and determination of their susceptibility to antimicrobial peptides

AU - Liu, Shiqi

AU - Brul, Stanley

AU - Zaat, Sebastian A. J.

N1 - Funding Information: Funding: This research in the labs of S.B. and S.A.J.Z. was supported by Chinese Scholarship Coun‐ cil grant (201904910554) awarded to S.L. Funding Information: Acknowledgments: We acknowledge Marco Lezzerini and Richard de Boer for providing help in qPCR experiments, and Gonzalo Congrains Sotomayor in FACS experiments. S.L. acknowledges the China Scholarship Council for her PhD scholarship. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2021/9/1

Y1 - 2021/9/1

N2 - Persister cells are growth‐arrested subpopulations that can survive possible fatal environments and revert to wild types after stress removal. Clinically, persistent pathogens play a key role in antibiotic therapy failure, as well as chronic, recurrent, and antibiotic‐resilient infections. In general, molecular and physiological research on persister cells formation and compounds against persister cells are much desired. In this study, we firstly demonstrated that the spore forming Grampositive model organism Bacillus subtilis can be used to generate persister cells during exposure to antimicrobial compounds. Interestingly, instead of exhibiting a unified antibiotic tolerance profile, different number of persister cells and spores were quantified in various stress conditions. qPCR results also indicated that differential stress responses are related to persister formation in various environmental conditions. We propose, for the first time to the best of our knowledge, an effective method to isolate B. subtilis persister cells from a population using fluorescence‐activated cell sorting (FACS), which makes analyzing persister populations feasible. Finally, we show that alphahelical cationic antimicrobial peptides SAAP‐148 and TC‐19, derived from human cathelicidin LL‐ 37 and human thrombocidin‐1, respectively, have high efficiency against both B. subtilis vegetative cells and persisters, causing membrane permeability and fluidity alteration. In addition, we confirm that in contrast to persister cells, dormant B. subtilis spores are not susceptible to the antimicrobial peptides.

AB - Persister cells are growth‐arrested subpopulations that can survive possible fatal environments and revert to wild types after stress removal. Clinically, persistent pathogens play a key role in antibiotic therapy failure, as well as chronic, recurrent, and antibiotic‐resilient infections. In general, molecular and physiological research on persister cells formation and compounds against persister cells are much desired. In this study, we firstly demonstrated that the spore forming Grampositive model organism Bacillus subtilis can be used to generate persister cells during exposure to antimicrobial compounds. Interestingly, instead of exhibiting a unified antibiotic tolerance profile, different number of persister cells and spores were quantified in various stress conditions. qPCR results also indicated that differential stress responses are related to persister formation in various environmental conditions. We propose, for the first time to the best of our knowledge, an effective method to isolate B. subtilis persister cells from a population using fluorescence‐activated cell sorting (FACS), which makes analyzing persister populations feasible. Finally, we show that alphahelical cationic antimicrobial peptides SAAP‐148 and TC‐19, derived from human cathelicidin LL‐ 37 and human thrombocidin‐1, respectively, have high efficiency against both B. subtilis vegetative cells and persisters, causing membrane permeability and fluidity alteration. In addition, we confirm that in contrast to persister cells, dormant B. subtilis spores are not susceptible to the antimicrobial peptides.

KW - Antimicrobial peptide

KW - Isolation

KW - Persisters

UR - http://www.scopus.com/inward/record.url?scp=85115052802&partnerID=8YFLogxK

U2 - 10.3390/ijms221810059

DO - 10.3390/ijms221810059

M3 - Article

C2 - 34576222

VL - 22

JO - International journal of molecular sciences

JF - International journal of molecular sciences

SN - 1661-6596

IS - 18

M1 - 10059

ER -