Culture-dependent and independent approaches for identifying novel halogenases encoded by Crambe crambe (marine sponge) microbiota

Sponges harbour microbial communities that contribute to the genetic and metabolic potential of their host. Among metabolites produced by sponge-associated microbial communities, halogenated compounds are of special interest because of their biotechnological potential. In this study, we have examined the diversity of the cultivable fraction of the marine demosponge Crambe crambe microbiota. Application of complementary cultivation methods yielded 107 bacterial isolates, some of which may be sponge-specific based on their phylogenetic analysis. Among these, Psychrobacter sp. was found to contain a putative halogenase gene. In addition to the culture-dependent approach for discovering halogenase genes, a cDNA library was constructed to determine the diversity of halogenase genes expressed in situ by the C. crambe microbiota. To this end, seventeen putative tryptophan halogenase cDNA sequences were identified, most of which were only remotely related to known halogenase genes, indicating the potential for novel bioactive compounds being produced by the C. crambe microbiota