Cytotoxic Compounds of Two Demosponges (Aplysina aerophoba and Spongia sp.) from the Aegean Sea

The class of demosponges is the biggest and most diverse of all described sponge species and it is reported to produce a plethora of chemically different metabolites with interesting biological activities. The focus of the present study was to investigate the chemical composition of two Mediterranean demosponges, targeting their brominated compounds and prenylated hydroquinones, compounds with interesting cytotoxic and anti-microbial properties. In order to gain a deeper insight into the chemical diversity of their metabolites and their activities, 20 pure secondary metabolites including new natural products were isolated from two different species (Aplysina aerophoba and Spongia sp.) using various chromatographic techniques. Their structures were confirmed by NMR and HRMS, revealing molecules with various chemical scaffolds, mainly prenylated hydroquinones from Spongia sp. and halogenated compounds from Aplysina aerophoba, including 5 novel natural products. The isolated compounds were investigated for their cytotoxic properties using 9 different cell lines, and especially one compound, 2,6-dibromo-4-hydroxy-4-methoxycarbonylmethylcyclohexa-2,5-dien-1-one showed good activities in all tested models.

Opisthobranch grazing results in mobilisation of spherulous cells and re-allocation of secondary metabolites in the sponge Aplysina aerophoba

Sponges thrive in marine benthic communities due to their specific and diverse chemical arsenal against predators and competitors. Yet, some animals specifically overcome these defences and use sponges as food and home. Most research on sponge chemical ecology has characterised crude extracts and investigated defences against generalist predators like fish. Consequently, we know little about chemical dynamics in the tissue and responses to specialist grazers. Here, we studied the response of the sponge Aplysina aerophoba to grazing by the opisthobranch Tylodina perversa, in comparison to mechanical damage, at the cellular (via microscopy) and chemical level (via matrix-assisted laser desorption/ionization imaging mass spectrometry, MALDI-imaging MS). We characterised the distribution of two major brominated alkaloids in A. aerophoba, aerophobin-2 and aeroplysinin-1, and identified a generalised wounding response that was similar in both wounding treatments: (i) brominated compound-carrying cells (spherulous cells) accumulated at the wound and (ii) secondary metabolites reallocated to the sponge surface. Upon mechanical damage, the wound turned dark due to oxidised compounds, causing T. perversa deterrence. During grazing, T. perversa’s way of feeding prevented oxidation. Thus, the sponge has not evolved a specific response to this specialist predator, but rather relies on rapid regeneration and flexible allocation of constitutive defences.

Grazing results in mobilization of spherulous cells and re-allocation of secondary metabolites to the surface in the sponge Aplysina aerophoba

On the sea floor, prey and predator commonly engage in a chemical warfare. Here, sponges thrive due to their specific and diverse chemical arsenal. Yet, some animals use these chemically-defended organisms as food and home. Most research on sponge chemical ecology has characterized crude extracts and investigated defences against generalist predators like fish. Consequently, we know little about intraindividual chemical dynamics and responses to specialist grazers. Here, we studied the response of the sponge Aplysina aerophoba to grazing by the opistobranch Tylodina perversa, in comparison to mechanical damage, at the cellular (via microscopy) and chemical level (via matrix-assisted laser desorption/ionization imaging mass spectrometry). We characterized the distribution of two major brominated compounds in A. aerophoba, aerophobin-2 and aeroplysinin-1, and identified a generalized wounding response that was similar in both wounding treatments: (i) brominated compound-carrying cells (spherulous cells) accumulated at the wound and (ii) secondary metabolites reallocated to the sponge surface. Upon mechanical damage, the wound turned dark due to oxidized compounds, causing T. perversa deterrence. During grazing, T. perversa’s way of feeding prevented oxidation. Thus, the sponge has not evolved a specific response to this specialist predator, but rather relies on rapid regeneration and flexible allocation of constitutive defences.

Antimicrobial activity investigation of natural bromotyrosine derivatives

Annotation. The relevance of the search for new natural antimicrobials is dictated by global spread of antimicrobial resistance. Studies of marine sponges of the class Demospongiae, including more than 7300 species, have become promising from the pharmacological point of view. Sponges produce a wide range of biologically active substances of different classes such as steroids, isoprene, alkaloids, terpenoids, etc., which can inhibit the growth of pathogenic microorganisms or kill them. The trial of brominated isoxazoline alkaloid derivates named bromotyrosines has been enough promising. The aim of our work was to determine the antimicrobial activity of aeroplysinin-1 against clinical antibiotic-resistant strains of gram-positive bacteria and fungi of the genus Candida. The aeroplysinin-1 substance was isolated by methanol extraction from desiccated specimens of sponge Aplysina aerophoba followed with its separation by column chromatography in the Extreme Biomimetics Laboratory of the Electronics and Sensor Materials Institute, Freiberg Mining Academy, UT (Germany). Well method was used to determine antimicrobial action of the tested substance against clinical strains of bacteria and fungi (S. aureus, C. acnes, C. albicans). Aeroplysinin-1 was found to demonstrate high antibacterial activity against clinical strains of the genera Cutibacterium, Staphylococcus. The inhibiting growth zones around the wells filled with 0.1% solution of aeroplysinin-1 in dimethyl sulfoxide had diameters of 35.4±3.2 mm, 32.1±2.8 mm and 26.5±2.5 mm for cutibacterium, enterococci, and staphylococci, respectively. It was revealed aeroplysinin-1 had no activity against fungi of the genus Candida. So, after a comprehensive study of pharmacological properties of this natural bromotyrosine compound, which is a secondary metabolite of marine sponges Aplysina aerophoba, it can be added to the list of alternative compounds to struggle multiresistent to modern antibiotics pathogens causing human diseases.

Diversity of tryptophan halogenases in sponges of the genus Aplysina

ABSTRACT Marine sponges are a prolific source of novel enzymes with promising biotechnological potential. Especially halogenases, which are key enzymes in the biosynthesis of brominated and chlorinated secondary metabolites, possess interesting properties towards the production of pharmaceuticals that are often halogenated. In this study we used a polymerase chain reaction (PCR)-based screening to simultaneously examine and compare the richness and diversity of putative tryptophan halogenase protein sequences and bacterial community structures of six Aplysina species from the Mediterranean and Caribbean seas. At the phylum level, bacterial community composition was similar amongst all investigated species and predominated by Actinobacteria, Chloroflexi, Cyanobacteria, Gemmatimonadetes, and Proteobacteria. We detected four phylogenetically diverse clades of putative tryptophan halogenase protein sequences, which were only distantly related to previously reported halogenases. The Mediterranean species Aplysina aerophoba harbored unique halogenase sequences, of which the most predominant was related to a sponge-associated Psychrobacter-derived sequence. In contrast, the Caribbean species shared numerous novel halogenase sequence variants and exhibited a highly similar bacterial community composition at the operational taxonomic unit (OTU) level. Correlations of relative abundances of halogenases with those of bacterial taxa suggest that prominent sponge symbiotic bacteria, including Chloroflexi and Actinobacteria, are putative producers of the detected enzymes and may thus contribute to the chemical defense of their host.