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

Abstract

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° 34′ 05″ N, 73° 44′ 17° 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 49.5 U mL−1 of protease and 3.67 U mL−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 700 U of penicillin. Cells were Gram-negative cocci, arranged in pairs, and 1-μm size in SEM micrograph. It possessed bacterioruberin with absorption at 387, 468, 492, and 523 nm. 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.