Isolation and culturing of protease- and lipase-producing Halococcus agarilyticus GUGFAWS-3 from marine Haliclona sp. inhabiting the rocky intertidal region of Anjuna in Goa, India

Three extremely halophilic bionts GUGFAWS-1, GUGFAWS-2, and GUGFAWS-3 were isolated from a marine, white sponge, attached to rocks, in the intertidal region of Anjuna, Goa, India (15 degrees 34 05 N, 73 degrees 44 17 degrees 40 E). Because the sponge had irregular tubes arranged in clusters, it was identified as Haliclona sp. All sponge bionts produced protease and lipase. GUGFAWS-1 and GUGFAWS-2 were euryhaline Eubacteria, growing from 0 to 30% NaCl concentration. The biont, GUGFAWS-3, was a haloarchaeon having glycerol diether moieties in its cells and showed R-O-R and long isoprenoid chains, in FTIR. The haloarchaeon, GUGFAWS-3, simultaneously produced extracellular 49.5UmL(-1) of protease and 3.67UmL(-1) of lipase, in the presence of 25% NaCl. It grew as dark orange-red colonies at 5-30% NaCl. Its growth was sensitive to bile salts and resistant to 700U of penicillin. Cells were Gram-negative cocci, arranged in pairs, and 1-m size in SEM micrograph. It possessed bacterioruberin with absorption at 387, 468, 492, and 523nm. The 16S rRNA gene sequence of GUGFAWS-3 was 99.1% similar to Halococcus agarilyticus 62E (T) of the family Halococcaceae of the domain Archaea. This study is the first evidence of retrieval and culturing of Halococcus agarilyticus strain GUGFAWS-3 (MF425611) from marine Haliclona sp. with ability to simultaneously produce protease and lipase extremozymes of ecological and biotechnological significance.