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
- 79 Downloads
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.
KeywordsMarine Haliclona sp. Haloarchaea Protease Lipase Extremozymes
S. K. Gaonkar gratefully acknowledges Goa University, research studentship award.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Aguiar R, Furtado I (1996) Growth of Halobacterium strain R3on sodium benzoate. In: RS Kahlon (ed) Perspectives in microbiology. Natl. Agri Tech Inf Centre India, pp 78–79Google Scholar
- Braganza JM, Furtado I (2009) Isolation of Haloarchaea from low-salinity coastal sediments and waters of Goa. Curr Sci 96:1182–1184Google Scholar
- Gaitonde D, Sarkar A, Kaisary S, Silva CD, Dias C, Rao DP, Ray D, Nagarajan R, De Sousa SN, Sarker S, Patil D (2006) Acetylcholinesterase activities in marine snail (Cronia contracta) as a biomarker of neurotoxic contaminants along the Goa coast, West coast of India. Ecotoxicol 15:353–358CrossRefGoogle Scholar
- Gaonkar SK, Furtado I (2018) Use of immobilized whole cells of Haloferax ATCC BAA 645 for treatment of dairy and fish waste effluents. IJPBS 8:16–21Google Scholar
- Grant RE (1841) Porifera. Pp 5–9, 310–313, pls II-IV. In: H Bailliere (ed) Outlines of comparative anatomy. 1 London, pp 1–656Google Scholar
- Grant WD, Kamekura M, McGenity TJ, Ventosa A (2001) Order I. Halobacteriales. In: Garrity GM (ed) Bergey’s manual of systematic bacteriology V.I, The Archaea and deeply branching and phototrophic bacteria, 2nd edn. Springer, New YorkGoogle Scholar
- Hammer Q, Harper DAT, Ryan PD (2001) PAST: paleontological statistics software package for education and data analysis. Palaeontol Electron 4(1):9ppGoogle Scholar
- Raghavan TM, Furtado I (2004) Occurrence of extremely haloarchaea in sediments from the continental shelf of west coast of India. Curr Sci 86:1065–1067Google Scholar
- Ross HNM, Collins MD, Tindall BJ, Grant WD (1981) A rapid procedure for the detection of archaebacterial lipids in halophilic bacteria. J Gen Microbiol 123:75–80Google Scholar
- Shi WL, Zhong CQ, Tang B, Shen P (2006) Purification and characterization of extracellular halophilic protease from haloarchaea Natrinema sp. R6-5. Acta Microbiol Sin 47(1):163–163Google Scholar
- Smibert RM, Krieg NR (1994) In: Gerhardt P (ed) Phenotypic characterization in: Methods for general and molecular bacteriology. American Society for Microbiology, Washington DC, pp 607–654Google Scholar
- Sneath PHA, Mair NS, Sharpe ME, Holt JG (1986) Bergey’s manual of systematic bacteriology, vol 2. Williams and Wilkins, BaltimoreGoogle Scholar
- Velho-Pereira S, Furtado I (2014) Retrieval of euryhaline eubacterial and haloarchaeal bionts from nine different benthic sponges: reflection of the bacteriological health of waters of Mandapam, India. Indian J Mar Sci 43:773–783Google Scholar
- Zhao GY, Chen XL, Zhao HL, Xie BB, Zhou BC, Zhang YZ (2008) Hydrolysis of insoluble collagen by deseasin MCP-03 from deep-sea Pseudoalteromonas sp. SM9913: collagenolytic characters, collagen binding ability of C-terminal polycystic kidney disease domain, and implication for its novel role in deep-sea sedimentary particulate organic nitrogen degradation. J Biol Chem 283:36100–36107CrossRefGoogle Scholar