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PreprintCited by 2Microbial Natural Products and BiosynthesisAntimicrobial Peptides and ActivitiesChemical Synthesis and Analysis

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Structure, biosynthesis, and bioactivity of nostolysamides

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Weir, Enleyona, Anterola, Ivan, van der Donk, Wilfred A.·2026·Biochemistry
DOI 10.64898/2026.01.31.703028

Preprint — published as 10.1021/acs.biochem.6c00103

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Abstract

A recent genome mining study identified class II lanthipeptides encoded in Nostoc punctiforme PCC 73102 that contain acyl groups conjugated to Lys side chains. The structure and bioactivity of these peptides, named nostolysamides, were not determined. In this study, we produced the nostolysamides by co-expression of the NpuA precursor peptide with an N-terminal SUMO tag with the class II lanthipeptide synthetase NpuM in Escherichia coli. All four lanthionine and methyllanthionine residues were shown to have the DL configuration by Marfey’s analysis. Tandem mass spectrometry and mutagenesis studies indicate an N-terminal non-overlapping methyllanthionine ring and three overlapping rings at the C-terminus for which the most likely ring pattern is proposed. The NpuM lanthipeptide synthetase is a member of the ProcM-clade and catalyzes ring formation with both C-to-N and N-to-C directionality. After removal of the leader peptide, the resulting lanthipeptide exhibits antibacterial as well as antifungal activity against Candida species by disrupting cell membranes. Antibacterial activity is shown not to involve lipid II. The biosynthetic gene cluster also encodes an acetyltransferase NpuN that transfers long chain acyl groups to the side chain of a Lys residue in position 1 of the precursor peptide. In vitro studies of NpuN show relatively broad substrate specificity with NpuN conjugating various acyl groups from acyl-CoA substrates to Lys1 in the nostolysamides. The acylation did not appreciably change the antifungal and antimicrobial activity of nostolysamide showing that it is not required for these activities.

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Metadata

Authors
Weir, E., Anterola, I., & van der Donk, W. A.
Affiliations
Howard Hughes Medical Institute (US)
Corresponding author
van der Donk, Wilfred A.
ORCID
Venue
Biochemistry
Publisher
openRxiv
Published
2026-01-31
Volume
65
Issue
9
Pages
1578-1591
ISSN
0006-2960 (ISSN-L)
DOI
10.64898/2026.01.31.703028
OpenAlex
W7155643082
Cited by
2
References
1
Citations by year
2026: 2
FWCI
8.43
Type
journal article
Language
en
Open access
Licence
Locations
  • Biochemistry (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion)
Funders
  • Howard Hughes Medical Institute
  • Roy J. Carver Charitable Trust
  • National Institutes of Health
  • National Institute of Allergy and Infectious Diseases (R01AI144967)
Topics
  • Life Sciences › Immunology and Microbiology › Applied Microbiology and Biotechnology › Microbial Natural Products and Biosynthesis
  • Life Sciences › Immunology and Microbiology › Microbiology › Antimicrobial Peptides and Activities
  • Physical Sciences › Chemistry › Organic Chemistry › Chemical Synthesis and Analysis
Keywords
Nostoc punctiforme, lanthionine, biosynthetic gene clusters, genome mining, antibacterial activity, antifungal activity, Candida, membrane damage, ring structure

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CoChat. (2026). Citation check: Structure, biosynthesis, and bioactivity of nostolysamides. https://app.cochat.ai/papers/10.64898/2026.01.31.703028
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Weir, E., Anterola, I., & van der Donk, W. A. (2026). Structure, biosynthesis, and bioactivity of nostolysamides. Biochemistry. https://doi.org/10.64898/2026.01.31.703028