TY - JOUR
T1 - In-depth phospholipid profiling of plant-pathogenic bacteria after treatment with antimicrobial random peptide mixtures
AU - Rudt, Edward
AU - Faist, Christian
AU - Schwantes, Vera
AU - Wiedmaier-Czerny, Nina
AU - Lehnert, Katja
AU - Topman-Rakover, Shiri
AU - Brill, Aya
AU - Burdman, Saul
AU - Hayouka, Zvi
AU - Vetter, Walter
AU - Hayen, Heiko
N1 - Publisher Copyright:
© 2025 The Authors
PY - 2025/3/8
Y1 - 2025/3/8
N2 - Background: The ability of plant-pathogenic bacteria to develop antimicrobial resistance against crop protection products represents a significant challenge. An alternative to conventional crop protecting products could be random peptide mixtures (RPMs), which potentially target the phospholipid-containing cell membrane. The randomized arrangement of the peptides minimizes the risk of bacterial resistance developing against the RPMs. However, not all plant-pathogenic bacteria exhibited growth inhibition after RPM treatment. Our prior studies revealed correlations between bacterial growth inhibition and changes in the fatty acid pattern following treatment. However, additional data on the intact phospholipid composition are essential to further understand and improve novel RPMs. Results: Accordingly, we developed an analytical setup for in-depth bacterial lipid membrane characterization based on two complementary methods in conjunction with chemometric data evaluation to study the impact of RPM treatment on phospholipid class and species level. An efficient phospholipid class quantitation using hydrophilic interaction liquid chromatography (HILIC)-based lipid class separation with uniform charged aerosol detection (CAD) revealed distinct differences in the class composition of six plant-pathogenic bacteria. Moreover, branched-chain fatty acid (BCFA)-comprising phospholipid profiling via liquid chromatography-tandem mass spectrometry (LC-MS/MS) provided additional lipid species information to classify the investigated bacteria based on the number of bound BCFA. The combination of these techniques served for a comprehensive characterization of the bacterial membrane adaptation to the RPM treatment, which showed some correlations with the inhibitory effects of the RPMs. Significance: In this proof-of-concept study, HILIC-CAD phospholipid quantitation and BCFA-comprising phospholipid profiling were introduced as complementary techniques for in-depth characterization of bacterial cell membranes as well as membrane adaptations at both phospholipid class and species level. Our developed analytical setup may facilitate future studies targeting in-depth characterization of bacterial lipid membranes.
AB - Background: The ability of plant-pathogenic bacteria to develop antimicrobial resistance against crop protection products represents a significant challenge. An alternative to conventional crop protecting products could be random peptide mixtures (RPMs), which potentially target the phospholipid-containing cell membrane. The randomized arrangement of the peptides minimizes the risk of bacterial resistance developing against the RPMs. However, not all plant-pathogenic bacteria exhibited growth inhibition after RPM treatment. Our prior studies revealed correlations between bacterial growth inhibition and changes in the fatty acid pattern following treatment. However, additional data on the intact phospholipid composition are essential to further understand and improve novel RPMs. Results: Accordingly, we developed an analytical setup for in-depth bacterial lipid membrane characterization based on two complementary methods in conjunction with chemometric data evaluation to study the impact of RPM treatment on phospholipid class and species level. An efficient phospholipid class quantitation using hydrophilic interaction liquid chromatography (HILIC)-based lipid class separation with uniform charged aerosol detection (CAD) revealed distinct differences in the class composition of six plant-pathogenic bacteria. Moreover, branched-chain fatty acid (BCFA)-comprising phospholipid profiling via liquid chromatography-tandem mass spectrometry (LC-MS/MS) provided additional lipid species information to classify the investigated bacteria based on the number of bound BCFA. The combination of these techniques served for a comprehensive characterization of the bacterial membrane adaptation to the RPM treatment, which showed some correlations with the inhibitory effects of the RPMs. Significance: In this proof-of-concept study, HILIC-CAD phospholipid quantitation and BCFA-comprising phospholipid profiling were introduced as complementary techniques for in-depth characterization of bacterial cell membranes as well as membrane adaptations at both phospholipid class and species level. Our developed analytical setup may facilitate future studies targeting in-depth characterization of bacterial lipid membranes.
KW - Branched-chain fatty acid
KW - HILIC-CAD
KW - LC-MS/MS
KW - Lipidomics
KW - Phospholipid
KW - Plant-pathogenic bacteria
KW - RPM
UR - http://www.scopus.com/inward/record.url?scp=85215857451&partnerID=8YFLogxK
U2 - 10.1016/j.aca.2025.343680
DO - 10.1016/j.aca.2025.343680
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AN - SCOPUS:85215857451
SN - 0003-2670
VL - 1342
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
M1 - 343680
ER -