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Identification and Biotechnological Potential of New Alcanivorax Species from Eutrophic Sea Pit in the Adriatic Sea

Microorganisms adapted to harsh marine habitats are considered as untapped sources of novel enzymes functional under diverse conditions and with unique biocatalytic potential for numerous industrial applications. A microbial diversity study was performed exploring undersea pit piled with ~15m high P. oceanica waste in central Eastern Adriatic. The entrance to the pit is surrounded by meadows of P. oceanica and is located 7m under the sea surface and extends to 32m deep. Out of 298 cultivable isolates, 67 isolates exhibited 99 % identity of 16S rRNA gene to Gammaproteobacteria Alcanivorax borkumensis. In order to get more precise taxonomical information, RpoB molecular marker was used on 4 Alcanivorax isolates. The sequencing results showed only 90% similarity to A. borkumensis indicating that we isolated so far unreported species. This genus is known to thrive on petroleum and is usually found near the sea surface or most abundantly in oil spills. While our isolates had slow, transparent growth on different types of common agar including marine agar, they thrived on media supplemented with Tween20 and Tween80, showing strong esterase and lipase activity. When media contained 1 % oil, this microorganism can tolerate up 10 % salt, and form specific mold-like hydrophobic clumps at the media surface. Additionally, this bacteria has the ability to grow on olive oil as the only source of nutrients. Confocal time-lapse microscopy has shown that this bacteria can colonize oil droplets more rapidly than previously described species. Different enzymatic assays were performed and revealed capacity of this bacterium to degrade various substrates. In addition whole genome sequencing of this bacteria is ongoing and will revealed genomic potential of this bacteria for bioremediation.

microbial diversity, bioremediation, Alcanivorax

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