Biosynthesis and Characterization of Novel Antimicrobial Peptides from Marine Bacteria
DOI:
https://doi.org/10.5281/zenodo.22688063Keywords:
antimicrobial peptides, marine bacteria, Pseudoalteromonas, genome mining, ribosomally synthesised peptides, multidrug-resistant pathogensAbstract
The rise of multidrug-resistant bacterial pathogens has renewed interest in under-explored environmental niches
as sources of novel antimicrobial chemistry, and marine sediments, with their steep gradients of pressure, salinity
and nutrient availability, host bacterial lineages whose secondary metabolite repertoires remain comparatively
poorly catalogued relative to soil-derived Streptomyces and Bacillus. We screened 46 culturable bacterial isolates
from intertidal and subtidal sediment samples collected off the North Sea coast for antagonistic activity against a
panel of six indicator pathogens including methicillin-resistant Staphylococcus aureus and carbapenem-resistant
Klebsiella pneumoniae, identifying nine isolates with reproducible zone-of-inhibition activity. Genome sequencing
and antiSMASH-guided mining of the three most active isolates, all belonging to the genus Pseudoalteromonas,
identified a previously uncharacterised ribosomally synthesised and post-translationally modified peptide (RiPP)
biosynthetic gene cluster present in all three strains. Heterologous expression of the cluster in Escherichia coli,
followed by reversed-phase HPLC purification and mass-spectrometry-guided structure elucidation, yielded a
novel 27-residue lanthipeptide, which we have named pseudoalterin, containing two lanthionine bridges
introduced by a dedicated LanM-type modification enzyme. Pseudoalterin showed potent activity against
Gram-positive indicator strains, including MRSA (minimum inhibitory concentration 2 microg/mL) and
vancomycin-resistant Enterococcus faecium (4 microg/mL), with negligible activity against the Gram-negative
panel and low haemolytic activity against human erythrocytes at concentrations up to 64 microg/mL. Mechanistic
assays indicated that pseudoalterin acts primarily through binding to the lipid II cell-wall precursor and consequent
disruption of peptidoglycan synthesis, a mechanism shared with several clinically used lantibiotics. These findings
identify marine sediment Pseudoalteromonas as a source of chemically novel, mechanistically validated
antimicrobial lanthipeptides warranting further preclinical characterisation.
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