Application of HPCCC Combined with Polymeric Resins and HPLC for the Separation of Cyclic Lipopeptides Muscotoxins A–C and Their Antimicrobial Activity

Abstract

Muscotoxins are cyanobacterial cyclic lipopeptides with potential applications in biomedicine and biotechnology. In this study, Desmonostoc muscorum CCALA125 strain extracts were enriched by polymeric resin treatment, and subjected to HPCCC affording three cyclic lipopeptides (1–3), which were further repurified by semi-preparative HPLC, affording 1, 2, and 3, with a purity of 86%, 92%, and 90%, respectively. The chemical identities of 2–3 were determined as muscotoxins A and B, respectively, by comparison with previously reported ESI-HRMS/MS data, whereas 1 was determined as a novel muscotoxin variant (muscotoxin C) using NMR and ESI-HRMS/MS data. Owing to the high yield (50 mg), compound 2 was broadly screened for its antimicrobial potential exhibiting a strong antifungal activity against Alternaria alternata, Monographella cucumerina, and Aspergillus fumigatus, with minimum inhibitory concentration (MIC) values of 0.58, 2.34, and 2.34 µg/mL; respectively, and weak antibacterial activity against Bacillus subtilis with a MIC value of 37.5 µg/mL. Compounds 1 and 3 were tested only against the plant pathogenic fungus Sclerotinia sclerotiorum due to their low yield, displaying a moderate antifungal activity. The developed chromatographic method proved to be an efficient tool for obtaining muscotoxins with potent antifungal properties.

Chemical diversity and cellular effects of antifungal cyclic lipopeptides from cyanobacteria

“…Cyanobacteria produce additional cyclic lipopeptides with long hydrocarbon chains with antifungal and/or antiproliferative activity (Niedermeyer, 2015), that do not fall into any of these four structural classes. Muscotoxins A–C are cyclic lipopeptides consisting of a long fatty acid side chain and 14 amino acid residues produced by Desmonostoc muscorum (Cheel et al, 2018; Tomek et al, 2015). Muscotoxins A and B manifest quite prominent inhibition effects that were reached against the plant pathogens Alternaria alternata , Monographella cucumerina , and Aspergillus fumigatus , with minimum inhibitory concentration values of 0.58, 2.34, and 2.34 μg mL −1 (Cheel et al, 2018).…”

“…Muscotoxins A–C are cyclic lipopeptides consisting of a long fatty acid side chain and 14 amino acid residues produced by Desmonostoc muscorum (Cheel et al, 2018; Tomek et al, 2015). Muscotoxins A and B manifest quite prominent inhibition effects that were reached against the plant pathogens Alternaria alternata , Monographella cucumerina , and Aspergillus fumigatus , with minimum inhibitory concentration values of 0.58, 2.34, and 2.34 μg mL −1 (Cheel et al, 2018). Muscotoxins are thought to permeabilize phospholipid membranes by reducing their fluidity (Cheel et al, 2018; Tomek et al, 2015).…”

“…Muscotoxins A and B manifest quite prominent inhibition effects that were reached against the plant pathogens Alternaria alternata , Monographella cucumerina , and Aspergillus fumigatus , with minimum inhibitory concentration values of 0.58, 2.34, and 2.34 μg mL −1 (Cheel et al, 2018). Muscotoxins are thought to permeabilize phospholipid membranes by reducing their fluidity (Cheel et al, 2018; Tomek et al, 2015). Nostofungicidine is an antifungal cyclic lipopeptide isolated from Nostoc commune containing a long fatty acid side chain and eight amino acids residues (Kajiyama et al, 1998).…”

Production of Fucoxanthin from Phaeodactylum tricornutum Using High Performance Countercurrent Chromatography Retaining Its FOXO3 Nuclear Translocation-Inducing Effect

“…Given that this technology lacks solid support, it has many advantages over the solid chromatographic techniques including large sample loading capacity, low risk of irreversible adsorption and sample denaturation, high sample recovery, low consumption of solvents and great operational versatility, since the roles of mobile and stationary phases can be exchanged during the chromatographic operation [ 40 , 41 , 42 ]. The capacity of countercurrent chromatography for obtaining valuable compounds from microalgae biomass has been widely demonstrated [ 42 , 43 , 44 , 45 , 46 , 47 , 48 ] and its application at industrial level is already a reality [ 49 ]. High-speed countercurrent chromatography (HSCCC) has been used for the isolation of fucoxanthin from edible brown macroalgae species such as Laminaria japonica , Undaria pinnatifida and Sargassum fusiforme [ 50 ] applying a single sample injection method.…”

Fast Isolation of Flavonoids from the Endemic Species Nolana ramosissima I.M. Johnst and Its Endothelium-Independent Relaxation Effect in Rat Aorta

“…This methodology was broadly used to separate the flavonoids from plants and fruits [10][11][12][13]. HSCCC offer important advantages in separation of natural products: lower consumption of solvents, use of green chemistry solvents, such as water and ethyl acetate, no absorption on solid surfaces such as conventional column chromatography, very higher amounts of processing sample, introduction of crude extracts, and full recovery of natural products [14][15][16][17][18]. In this work we have applied this technique for the fast detection of flavonoids, from the methanolic extract of N. ramosissima for the testing of their relaxation activity in rat aorta.…”