scholarly journals Discovery of the Class I Antimicrobial Lasso Peptide Arcumycin

2021 ◽  
Author(s):  
Lydia Stariha ◽  
Dewey G. McCafferty
Keyword(s):  
Micrococcus Luteus ◽  
Genome Mining ◽  
Antibiotic Activity ◽  
Peptide Sequence ◽  
Class I ◽  
Gram Positive ◽  
Conformationally Constrained ◽  
Lasso Peptide ◽  
Lasso Peptides ◽  
Lipid Ii

<p>Lasso peptides are a structurally diverse superfamily of</p><p>conformationally-constrained peptide natural products, of which a</p><p>subset exhibits broad antimicrobial activity. Although advances in</p><p>bioinformatics have increased our knowledge of strains harboring</p><p>the biosynthetic machinery for lasso peptide production, relating</p><p>peptide sequence to bioactivity remains a continuous challenge.</p><p>Towards this end, a structure-driven genome mining investigation</p><p>of Actinobacteria-produced antimicrobial lasso peptides was</p><p>performed to correlate predicted primary structure with antibiotic</p><p>activity. Bioinformatic evaluation revealed eight putative novel</p><p>class I lasso peptide sequences. This subset is predicted to</p><p>possess antibiotic activity as characterized members of this class</p><p>have both broad spectrum and potent activity against Gram positive</p><p>strains. Fermentation of one of these hits, Streptomyces</p><p>NRRL F-5639, resulted in the production of a novel class I lasso</p><p>peptide, arcumycin, named for the Latin word for bow or arch,</p><p>arcum. Arcumycin exhibited antibiotic activity against Gram positive</p><p>bacteria including <i>Bacillus subtilis</i> (4 μg/mL),</p><p><i>Staphylococcus aureus </i>(8 μg/mL), and <i>Micrococcus luteus</i> (8</p><p>μg/mL). Arcumycin treatment of <i>B. subtilis</i> liaI-β-gal promoter</p><p>fusion reporter strain resulted in upregulation of the system liaRS</p><p>by the promoter liaI, indicating arcumycin interferes with lipid II</p><p>biosynthesis. Cumulatively, the results illustrate the relationship</p><p>between phylogenetically related lasso peptides and their</p><p>bioactivity as validated through the isolation, structural</p><p>determination, and evaluation of bioactivity of the novel class I</p><p>antimicrobial lasso peptide arcumycin.</p>

scholarly journals Discovery of the Class I Antimicrobial Lasso Peptide Arcumycin

2021 ◽  
Author(s):  
Lydia Stariha ◽  
Dewey G. McCafferty
Keyword(s):  
Micrococcus Luteus ◽  
Genome Mining ◽  
Antibiotic Activity ◽  
Peptide Sequence ◽  
Class I ◽  
Gram Positive ◽  
Conformationally Constrained ◽  
Lasso Peptide ◽  
Lasso Peptides ◽  
Lipid Ii

<p>Lasso peptides are a structurally diverse superfamily of</p><p>conformationally-constrained peptide natural products, of which a</p><p>subset exhibits broad antimicrobial activity. Although advances in</p><p>bioinformatics have increased our knowledge of strains harboring</p><p>the biosynthetic machinery for lasso peptide production, relating</p><p>peptide sequence to bioactivity remains a continuous challenge.</p><p>Towards this end, a structure-driven genome mining investigation</p><p>of Actinobacteria-produced antimicrobial lasso peptides was</p><p>performed to correlate predicted primary structure with antibiotic</p><p>activity. Bioinformatic evaluation revealed eight putative novel</p><p>class I lasso peptide sequences. This subset is predicted to</p><p>possess antibiotic activity as characterized members of this class</p><p>have both broad spectrum and potent activity against Gram positive</p><p>strains. Fermentation of one of these hits, Streptomyces</p><p>NRRL F-5639, resulted in the production of a novel class I lasso</p><p>peptide, arcumycin, named for the Latin word for bow or arch,</p><p>arcum. Arcumycin exhibited antibiotic activity against Gram positive</p><p>bacteria including <i>Bacillus subtilis</i> (4 μg/mL),</p><p><i>Staphylococcus aureus </i>(8 μg/mL), and <i>Micrococcus luteus</i> (8</p><p>μg/mL). Arcumycin treatment of <i>B. subtilis</i> liaI-β-gal promoter</p><p>fusion reporter strain resulted in upregulation of the system liaRS</p><p>by the promoter liaI, indicating arcumycin interferes with lipid II</p><p>biosynthesis. Cumulatively, the results illustrate the relationship</p><p>between phylogenetically related lasso peptides and their</p><p>bioactivity as validated through the isolation, structural</p><p>determination, and evaluation of bioactivity of the novel class I</p><p>antimicrobial lasso peptide arcumycin.</p>

scholarly journals Sarconesin II, a New Antimicrobial Peptide Isolated from Sarconesiopsis magellanica Excretions and Secretions

Molecules ◽  
2019 ◽  
Vol 24 (11) ◽  
pp. 2077 ◽  
Author(s):  
Andrea Díaz-Roa ◽  
Abraham Espinoza-Culupú ◽  
Orlando Torres-García ◽  
Monamaris M. Borges ◽  
Ivan N. Avino ◽  
...  
Keyword(s):  
Micrococcus Luteus ◽  
Multidrug Resistant ◽  
Peptide Sequence ◽  
Resistant Bacteria ◽  
Gram Positive ◽  
Antimicrobial Drugs ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Conserved Domain ◽  
Health Institutions

Antibiotic resistance is at dangerous levels and increasing worldwide. The search for new antimicrobial drugs to counteract this problem is a priority for health institutions and organizations, both globally and in individual countries. Sarconesiopsis magellanica blowfly larval excretions and secretions (ES) are an important source for isolating antimicrobial peptides (AMPs). This study aims to identify and characterize a new S. magellanica AMP. RP-HPLC was used to fractionate ES, using C18 columns, and their antimicrobial activity was evaluated. The peptide sequence of the fraction collected at 43.7 min was determined by mass spectrometry (MS). Fluorescence and electronic microscopy were used to evaluate the mechanism of action. Toxicity was tested on HeLa cells and human erythrocytes; physicochemical properties were evaluated. The molecule in the ES was characterized as sarconesin II and it showed activity against Gram-negative (Escherichia coli MG1655, Pseudomonas aeruginosa ATCC 27853, P. aeruginosa PA14) and Gram-positive (Staphylococcus aureus ATCC 29213, Micrococcus luteus A270) bacteria. The lowest minimum inhibitory concentration obtained was 1.9 μM for M. luteus A270; the AMP had no toxicity in any cells tested here and its action in bacterial membrane and DNA was confirmed. Sarconesin II was documented as a conserved domain of the ATP synthase protein belonging to the Fli-1 superfamily. The data reported here indicated that peptides could be alternative therapeutic candidates for use in infections against Gram-negative and Gram-positive bacteria and eventually as a new resource of compounds for combating multidrug-resistant bacteria.

The Lasso Peptide Siamycin-I Targets Lipid II at the Gram-Positive Cell Surface

2019 ◽  
Vol 14 (5) ◽  
pp. 966-974 ◽  
Author(s):  
Stephanie Tan ◽  
Kevin C. Ludwig ◽  
Anna Müller ◽  
Tanja Schneider ◽  
Justin R. Nodwell
Keyword(s):  
Cell Surface ◽  
Gram Positive ◽  
Lasso Peptide ◽  
Lipid Ii

scholarly journals Precursor-centric genome-mining approach for lasso peptide discovery

2012 ◽  
Vol 109 (38) ◽  
pp. 15223-15228 ◽  
Author(s):  
Mikhail O. Maksimov ◽  
István Pelczer ◽  
A. James Link
Keyword(s):  
Solution Structure ◽  
Caulobacter Crescentus ◽  
Genome Mining ◽  
Gene Clusters ◽  
Bacterial Phyla ◽  
Lasso Peptide ◽  
Lasso Peptides ◽  
Genes Encoding ◽  
Peptide Gene ◽  
Conserved Amino Acids

Lasso peptides are a class of ribosomally synthesized posttranslationally modified natural products found in bacteria. Currently known lasso peptides have a diverse set of pharmacologically relevant activities, including inhibition of bacterial growth, receptor antagonism, and enzyme inhibition. The biosynthesis of lasso peptides is specified by a cluster of three genes encoding a precursor protein and two enzymes. Here we develop a unique genome-mining algorithm to identify lasso peptide gene clusters in prokaryotes. Our approach involves pattern matching to a small number of conserved amino acids in precursor proteins, and thus allows for a more global survey of lasso peptide gene clusters than does homology-based genome mining. Of more than 3,000 currently sequenced prokaryotic genomes, we found 76 organisms that are putative lasso peptide producers. These organisms span nine bacterial phyla and an archaeal phylum. To provide validation of the genome-mining method, we focused on a single lasso peptide predicted to be produced by the freshwater bacterium Asticcacaulis excentricus. Heterologous expression of an engineered, minimal gene cluster in Escherichia coli led to the production of a unique lasso peptide, astexin-1. At 23 aa, astexin-1 is the largest lasso peptide isolated to date. It is also highly polar, in contrast to many lasso peptides that are primarily hydrophobic. Astexin-1 has modest antimicrobial activity against its phylogenetic relative Caulobacter crescentus. The solution structure of astexin-1 was determined revealing a unique topology that is stabilized by hydrogen bonding between segments of the peptide.

scholarly journals Cell-Free Biosynthesis to Evaluate Lasso Peptide Formation and Enzyme-Substrate Tolerance

2020 ◽  
Author(s):  
Yuanyuan Si ◽  
Ashley M. Kretsch ◽  
Laura M. Daigh ◽  
Mark J. Burk ◽  
Douglas A. Mitchell
Keyword(s):  
Genome Mining ◽  
Efficient Production ◽  
Surface Receptors ◽  
Lasso Peptide ◽  
Anticancer Properties ◽  
Lasso Peptides ◽  
Isolation And Characterization ◽  
Precursor Peptide ◽  
A Cell ◽  
Substrate Tolerance

AbstractLasso peptides are ribosomally synthesized and post-translationally modified peptide (RiPP) natural products that display a unique lariat-like structure. Owing to a rigid topology, lasso peptides are unusually stable towards heat and proteolytic degradation. Some lasso peptides have been shown to bind human cell-surface receptors and exhibit anticancer properties, while others display antibacterial or antiviral activities. Known lasso peptides are produced by bacteria and genome-mining studies indicate that lasso peptides are a relatively prevalent RiPP class; however, the discovery, isolation, and characterization of lasso peptides are constrained by the lack of an efficient production system. In this study, we employ a cell-free biosynthesis (CFB) strategy to address the longstanding challenges associated with lasso peptide production. We report the successful formation of a diverse array of lasso peptides that include known examples as well as a new predicted lasso peptide from Thermobifida halotolerans. We further demonstrate the utility of CFB to rapidly generate and characterize multisite precursor peptide variants in order to evaluate the substrate tolerance of the biosynthetic pathway. We show that the lasso-forming cyclase from the fusilassin pathway can produce millions of sequence-diverse lasso peptides via CFB with an extraordinary level of sequence permissiveness within the ring region of the lasso peptide. These data lay a firm foundation for the creation of large lasso peptide libraries using CFB to identify new variants with unique properties.

scholarly journals Cellulonodin-2 and Lihuanodin: Lasso Peptides with an Aspartimide Post-Translational Modification

2021 ◽  
Author(s):  
Li Cao ◽  
Moshe Beiser ◽  
Joseph D Koos ◽  
Margarita Orlova ◽  
Hader E Elashal ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Genome Mining ◽  
Gene Clusters ◽  
Post Translational Modification ◽  
Protein Repair ◽  
Lasso Peptide ◽  
Lasso Peptides ◽  
Modified Peptides ◽  
Asparagine Deamidation

Lasso peptides are a family of ribosomally synthesized and post-translationally modified peptides (RiPPs) defined by their threaded structure. Besides the class-defining isopeptide bond, other post-translational modifications (PTMs) that further tailor lasso peptides have been previously reported. Using genome mining tools, we identified a subset of lasso peptide biosynthetic gene clusters (BGCs) that are colocalized with protein L-isoaspartyl methyltransferase (PIMT) homologs. PIMTs have an important role in protein repair, restoring isoaspartate residues formed from asparagine deamidation to aspartate. Here we report a new function for PIMT enzymes in the post-translational modification of lasso peptides. The PIMTs associated with lasso peptide BGCs first methylate an L-aspartate sidechain found within the ring of the lasso peptide. The methyl ester is then converted into a stable aspartimide moiety, endowing the lasso peptide ring with rigidity relative to its unmodified counterpart. We describe the heterologous expression and structural characterization of two examples of aspartimide-modified lasso peptides from thermophilic Gram-positive bacteria. The lasso peptide cellulonodin-2 is encoded in the genome of actinobacterium Thermobifida cellulosilytica, while lihuanodin is encoded in the genome of firmicute Lihuaxuella thermophila. Additional genome mining revealed PIMT-containing lasso peptide BGCs in 48 organisms. In addition to heterologous expression, we have reconstituted PIMT-mediated aspartimide formation in vitro, showing that lasso peptide-associated PIMTs transfer methyl groups very rapidly as compared to canonical PIMTs. Furthermore, in stark contrast to other characterized lasso peptide PTMs, the methyltransferase functions only on lassoed substrates.

scholarly journals Angucycline-like Aromatic Polyketide from a Novel Streptomyces Species Reveals Freshwater Snail Physa acuta as Underexplored Reservoir for Antibiotic-Producing Actinomycetes

Antibiotics ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 22
Author(s):  
Nasim Safaei ◽  
Yvonne Mast ◽  
Michael Steinert ◽  
Katharina Huber ◽  
Boyke Bunk ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Genome Mining ◽  
Freshwater Snail ◽  
Whole Genome Sequence ◽  
Phylogenomic Analysis ◽  
Ionization Mass ◽  
Gram Positive ◽  
Streptomyces Species ◽  
Genus Streptomyces ◽  
Physa Acuta

Antibiotic producers have mainly been isolated from soil, which often has led to the rediscovery of known compounds. In this study, we identified the freshwater snail Physa acuta as an unexplored source for new antibiotic producers. The bacterial diversity associated with the snail was characterized by a metagenomic approach using cultivation-independent high-throughput sequencing. Although Actinobacteria represented only 2% of the bacterial community, the focus was laid on the isolation of the genus Streptomyces due to its potential to produce antibiotics. Three Streptomyces strains (7NS1, 7NS2 and 7NS3) were isolated from P. acuta, and the antimicrobial activity of the crude extracts were tested against a selection of Gram-positive and Gram-negative bacteria and fungi. 7NS3 showed the strongest activity against Gram-positive bacteria and, thus, was selected for genome sequencing and a phylogenomic analysis. 7NS3 represents a novel Streptomyces species, which was deposited as Streptomyces sp. DSM 110735 at the Leibniz Institute-German Collection of Microorganisms and Cell Cultures (DSMZ). Bioassay-guided high-performance liquid chromatography (HPLC) and high-resolution electrospray ionization-mass spectrometry (HR-ESI-MS) analyses of crude extract fractions resulted in the detection of four compounds, one of which matched the compound characteristics of emycin A, an angucycline-like aromatic polyketide. Genome mining studies based on the whole-genome sequence of 7NS3 resulted in the identification of a gene cluster potentially coding for emycin A biosynthesis. Our study demonstrates that freshwater snails like P. acuta can represent promising reservoirs for the isolation of new antibiotic-producing actinobacterial species.

scholarly journals Gram‐Positive and Gram‐Negative Antibiotic Activity of Asymmetric and Monomeric Robenidine Analogues

ChemMedChem ◽  
2018 ◽  
Vol 13 (23) ◽  
pp. 2573-2580 ◽  
Author(s):  
Cecilia C. Russell ◽  
Andrew Stevens ◽  
Hongfei Pi ◽  
Manouchehr Khazandi ◽  
Abiodun D. Ogunniyi ◽  
...  
Keyword(s):  
Antibiotic Activity ◽  
Gram Positive ◽  
Gram Negative

scholarly journals Proliferation of Antibiotic-Producing Bacteria and Concomitant Antibiotic Production as the Basis for the Antibiotic Activity of Jordan's Red Soils

2009 ◽  
Vol 75 (9) ◽  
pp. 2735-2741 ◽  
Author(s):  
Joseph O. Falkinham ◽  
Thomas E. Wall ◽  
Justin R. Tanner ◽  
Khaled Tawaha ◽  
Feras Q. Alali ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Activity ◽  
Incubation Period ◽  
Micrococcus Luteus ◽  
Antibiotic Production ◽  
Antibiotic Activity ◽  
Skin Infections ◽  
High Antimicrobial Activity ◽  
Methanol Extracts ◽  
Red Soils

ABSTRACT Anecdotes, both historical and recent, recount the curing of skin infections, including diaper rash, by using red soils from the Hashemite Kingdom of Jordan. Following inoculation of red soils isolated from geographically separate areas of Jordan, Micrococcus luteus and Staphylococcus aureus were rapidly killed. Over the 3-week incubation period, the number of specific types of antibiotic-producing bacteria increased, and high antimicrobial activity (MIC, ∼10 μg/ml) was observed in methanol extracts of the inoculated red soils. Antibiotic-producing microorganisms whose numbers increased during incubation included actinomycetes, Lysobacter spp., and Bacillus spp. The actinomycetes produced actinomycin C2 and actinomycin C3. No myxobacteria or lytic bacteriophages with activity against either M. luteus or S. aureus were detected in either soil before or after inoculation and incubation. Although protozoa and amoebae were detected in the soils, the numbers were low and did not increase over the incubation period. These results suggest that the antibiotic activity of Jordan's red soils is due to the proliferation of antibiotic-producing bacteria.

scholarly journals Put a Bow on It: Knotted Antibiotics Take Center Stage

Antibiotics ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 117 ◽  
Author(s):  
Stephanie Tan ◽  
Gaelen Moore ◽  
Justin Nodwell
Keyword(s):  
Current Knowledge ◽  
Chemical Diversity ◽  
Peptide Antibiotics ◽  
Linear Peptide ◽  
Peptidoglycan Biosynthesis ◽  
Lasso Peptides ◽  
Lipid Ii ◽  
Modified Peptides ◽  
Domains Of Life ◽  
Peptide Biosynthesis

Ribosomally-synthesized and post-translationally modified peptides (RiPPs) are a large class of natural products produced across all domains of life. The lasso peptides, a subclass of RiPPs with a lasso-like structure, are structurally and functionally unique compared to other known peptide antibiotics in that the linear peptide is literally “tied in a knot” during its post-translational maturation. This underexplored class of peptides brings chemical diversity and unique modes of action to the antibiotic space. To date, eight different lasso peptides have been shown to target three known molecular machines: RNA polymerase, the lipid II precursor in peptidoglycan biosynthesis, and the ClpC1 subunit of the Clp protease involved in protein homeostasis. Here, we discuss the current knowledge on lasso peptide biosynthesis as well as their antibiotic activity, molecular targets, and mechanisms of action.

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