Final Report: NOAA Ocean Exploration NOFO NA19OAR0110303 Bioprospecting for industrial enzymes and drug lead compounds in an ancient submarine forest

In this project, we addressed Objective 1 of the 2019 NOFO—discovering microorganisms, sponges, corals, and other organisms with biopharmaceutical or biotechnical potential—by mapping, documenting, describing, archiving, and exploring the biodiversity of a prehistoric submerged bald cypress forest off the coast of Alabama. Our primary focus was on wood-associated animal species and their microbiomes. We collected, identified, georeferenced, vouchered, and subsampled these animals, cultivated their associated bacterial symbionts, and sequenced the symbiont genomes and host metagenomes. We performed phylogenetic and bioinformatic analyses to identify these organisms and their associated antimicrobial compounds and hydrolytic enzymes capable of degrading the components of wood (lignocellulose). We then performed biochemical and growth inhibition assays to confirm the activities of these compounds. We discovered a new antibiotic with activity against Acinetobacter, an emerging multidrug-resistant pathogen of growing public health concern. We described and formally named one new bivalve genus and species and four new microbial species and submitted voucher specimens and isolates to appropriate public collections. We discovered and isolated at least 25 new bacterial species and hundreds of new bacterial strains and deposited them to the publicly accessible Ocean Genome Legacy biorepository. We showed that these bacteria, now available for further characterization, have great potential for drug and enzyme discovery. We determined the mechanism by which wood-boring bivalves (shipworms) produce and export cellulolytic enzymes and possibly antimicrobial compounds made by their symbiotic bacteria. Finally, we generated 3D maps of the site using photogrammetry and sidescan sonar. Our research demonstrated that marine wood deposits, like the Alabama undersea forest, are rich sources for discovering new animal and bacterial species and associated compounds of pharmaceutical and industrial value.