Evolutionary relevance of metabolite production in relation to marine sponge bacteria symbiont

Sponges are habitats for a diverse community of microorganisms. Sponges provide shelter, whereas microbes provide a complementary defensive mechanism. Here, a symbiotic bacterium, identified as Bacillus spp ., was isolated from a marine sponge following culture enrichment. Fermentation-assisted metabolomics using thin-layer chromatography (TLC) and gas chromatography-mass spectrometry (GC-MS) indicated that marine simulated nutrition and temperature was the optimum in metabolite production represented by the highest number of metabolites and the diverse chemical classes when compared to other culture media. Following large-scale culture in potato dextrose broth (PDB) and dereplication, compound M1 was isolated and identified as octadecyl-1-(2′,6′-di-tert-butyl-1′-hydroxyphenyl) propionate. M1 , at screening concentrations up to 10 mg/ml, showed no activity against prokaryotic bacteria including Staphylococcus aureus and Escherichia coli , while 1 mg/ml of M1 was sufficient to cause a significant killing effect on eukaryotic cells including Candida albicans , Candida auris , and Rhizopus delemar fungi and different mammalian cells. M1 exhibited MIC 50 0.97 ± 0.006 and 7.667 ± 0.079 mg/ml against C. albicans and C. auris , respectively. Like fatty acid esters, we hypothesize that M1 is stored in a less harmful form and upon pathogenic attack is hydrolyzed to a more active form as a defensive metabolite. Subsequently, [3-(3,5-di- tert -butyl-4-hydroxyphenyl)-propionic acid] (DTBPA), the hydrolysis product of M1 , exhibited ~ 8-fold and 18-fold more antifungal activity than M1 against C. albicans and C. auris , respectively. These findings indicated the selectivity of that compound as a defensive metabolite towards the eukaryotic cells particularly the fungi, a major infectious agent to sponges. Metabolomic-assisted fermentation can provide a significant understanding of a triple marine-evolved interaction. Key points • Bacillus species, closely related to uncultured Bacillus, is isolated from Gulf marine sponge • Metabolomic-assisted fermentations showed diverse metabolites • An ester with a killing effect against eukaryotes but not prokaryotes is isolated