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Article

High-Quality Nucleic Acid Isolation from Hard-to-Lyse Bacterial Strains Using PMAP-36, a Broad-Spectrum Antimicrobial Peptide

Department of Stem Cell and Regenerative Biotechnology, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 05029, Korea
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Author to whom correspondence should be addressed.
Authors contributed equally.
Academic Editor: Alina Maria Holban
Int. J. Mol. Sci. 2021, 22(8), 4149; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22084149
Received: 31 March 2021 / Revised: 13 April 2021 / Accepted: 14 April 2021 / Published: 16 April 2021
(This article belongs to the Special Issue Molecules and Nanoparticles to Fight Resistant Pathogens)
The efficiency of existing cell lysis methods to isolate nucleic acids from diverse bacteria varies depending on cell wall structures. This study tested a novel idea of using broad-spectrum antimicrobial peptides to improve the lytic efficiency of hard-to-lyse bacteria and characterized their differences. The lysis conditions of Staphylococcus aureus using recombinant porcine myeloid antimicrobial peptide 36 (PMAP-36), a broad-spectrum pig cathelicidin, was optimized, and RNA isolation was performed with cultured pellets of ten bacterial species using various membranolytic proteins. Additionally, three other antimicrobial peptides, protegrin-1 (PG-1), melittin, and nisin, were evaluated for their suitability as the membranolytic agents of bacteria. However, PMAP-36 use resulted in the most successful outcomes in RNA isolation from diverse bacterial species. The amount of total RNA obtained using PMAP-36 increased by ~2-fold compared to lysozyme in Salmonella typhimurium. Streptococci species were refractory to all lytic proteins tested, although the RNA yield from PMAP-36 treatment was slightly higher than that from other methods. PMAP-36 use produced high-quality RNA, and reverse transcription PCR showed the efficient amplification of the 16S rRNA gene from all tested strains. Additionally, the results of genomic DNA isolation were similar to those of RNA isolation. Thus, our findings present an additional option for high quality and unbiased nucleic acid isolation from microbiomes or challenging bacterial strains. View Full-Text
Keywords: antimicrobial peptides; PMAP-36; nucleic acids isolation antimicrobial peptides; PMAP-36; nucleic acids isolation
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MDPI and ACS Style

Cho, H.-s.; Choi, M.; Lee, Y.; Jeon, H.; Ahn, B.; Soundrarajan, N.; Hong, K.; Kim, J.-H.; Park, C. High-Quality Nucleic Acid Isolation from Hard-to-Lyse Bacterial Strains Using PMAP-36, a Broad-Spectrum Antimicrobial Peptide. Int. J. Mol. Sci. 2021, 22, 4149. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22084149

AMA Style

Cho H-s, Choi M, Lee Y, Jeon H, Ahn B, Soundrarajan N, Hong K, Kim J-H, Park C. High-Quality Nucleic Acid Isolation from Hard-to-Lyse Bacterial Strains Using PMAP-36, a Broad-Spectrum Antimicrobial Peptide. International Journal of Molecular Sciences. 2021; 22(8):4149. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22084149

Chicago/Turabian Style

Cho, Hye-sun; Choi, Munjeong; Lee, Yunjung; Jeon, Hyoim; Ahn, Byeongyong; Soundrarajan, Nagasundarapandian; Hong, Kwonho; Kim, Jin-Hoi; Park, Chankyu. 2021. "High-Quality Nucleic Acid Isolation from Hard-to-Lyse Bacterial Strains Using PMAP-36, a Broad-Spectrum Antimicrobial Peptide" Int. J. Mol. Sci. 22, no. 8: 4149. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22084149

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