Isolation, characterization and exploring biotechnological potential of halophilic archaea from salterns of western India
- 116 Downloads
Thirteen halophilic archaea were isolated from Kandla and Bhayander salt pans. These isolates were grouped into three different genera Halobacterium, Haloferax and Haloarcula based on morphological and biochemical characterization, polar lipid analysis, Amplified 16S rDNA restriction analysis (ARDRA) and 16S rDNA sequence analysis. Biochemical characterization suggested the ability of isolates to produce protease, amylase and poly-hydroxybutyrate (PHB) indicating their biotechnological potential. The isolates were further screened for the amount of extracellular protease produced. Halobacterium sp. SP1(1) showed significant protease production compared to other isolates. Protease producing ability of the isolate was influenced by several factors such as NaCl concentration, type of protein source, metal ions and surfactants, and presence of amino acid supplements in the production medium. Soybean flour, FeCl3 and dicotylsulfosuccinate were found to increase protease production by 2.36, 1.54 and 1.26 folds, respectively compared to production in basal medium. Effect of organic solvents used in paints (n-decane, n-undecane and n-dodecane) was also investigated on protease production by the isolate. Protease production by Halobacterium sp. SP1(1) was enhanced by 1.2 folds in presence of n-decane compared to control. Furthermore, the ability of isolate to hydrolyse fish protein was investigated using three different edible fishes (Pomfret, Flat fish and Seer fish) as sole protein source. Pomfret was found to be a good protein source for protease production by the isolate. These results revealed that Halobacterium sp. SP1(1) may have potential for paint-based antifouling coating preparations and fish sauce preparation by virtue of its extracellular protease.
KeywordsARDRA Fish protein Halobacterium Halophilic archaea Organic solvents Protease Salt pans
The work was supported by Council of Scientific and Industrial Research (CSIR) grant (90(0038)/04/EMR-II), New Delhi, India. The standard strain Halobacterium salinarum was gifted by Dr. H. M. Sonawat, Tata Institute of Fundamental Research, Mumbai, India. Heartiest thanks to Prof. A. J. Desai, M. S. University of Baroda, India, for providing lab facility and guidance.
Compliance with ethical standards
Conflict of interest
Authors report no conflict of interest.
- Akolkar AV (2009) Isolation and characterization of halophilic archaea: Production characterization and application of extracellular protease from Halobacterium SP SP1 1. Ph. D thesis, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, IndiaGoogle Scholar
- Akolkar AV, Durai D, Desai AJ (2010) Halobacterium sp. SP1 (1) as a starter culture for accelerating fish sauce fermentation. J Appl Microbiol 109:44–53Google Scholar
- Dave SR, Desai HB (2006) Microbial diversity at marine salterns near Bhavnagar, Gujarat, India. Curr Sci 90:497–500Google Scholar
- Dyall-Smith M (2008) The halohandbook: protocols for halobacterial genetics (Version 7 electronic publication)Google Scholar
- Grant WD, Larsen H (1989) Extremely halophilic archaeobacteria. In: Staley, Bryant, Pfennig and Holt (ed.) Bergy’s manual of systematic bacteriology, 1st edn. Vol 3, Williams & Wilkins Co, Baltimore, pp. 2216–2219Google Scholar
- Hampp N, Oesterhelt D (2008) Bacteriorhodopsin and its potential in technical applications. Protein Science Encyclopedia. Wiley-VCH Verlag GmbH & Co. KGaA WeinheimGoogle Scholar
- Kamekura M, Sen Y, Holmes ML, Dyall-Smith ML (1992) Molecular cloning and sequencing of the gene for a halophilic alkaline serine protease (halolysin) from an unidentified halophilic Archaea strain (172P1) and expression of the gene in Haloferax volcanii. J Bacteriol 174:736–742CrossRefGoogle Scholar
- Kerkar S (2004) Studies on bacteria of the dissimilatory reductive processes of the sulphur cycle from the salt pans of Goa. Ph.D, Thesis, Department of Marine Sciences, Goa University, Goa, IndiaGoogle Scholar
- Lillo JG, Rodriguez-Valera F (1990) Effect of culture conditions on poly-fi- hydroxybutyric acid production of Haloferax mediterranei. Appl Environ Microbiol 56:2517–2521Google Scholar
- Rohlf FJ (2009) NTSYSpc Numerical taxonomy and multivariate analysis system. Version 2.2. Getting Started Guide. Applied Biostatistics Inc., New YorkGoogle Scholar
- Thongthai C, Panbangred W, Choprasert C, Dhaveetiyanond S (1990) Protease activities in the traditional process of fish sauce fermentation. In: Reilly PJA, Parry RWH, Barile LE (eds) Post-harvest technology, preservation and quality of fish in Southeast Asia. Stockholm, International Foundation for Sciences, pp 61–65Google Scholar
- Tomlinson GA, Jahnke LL, Hochstein LI (1986) Halobacterium denitrificans sp nov, an extremely halophilic denitrifying bacterium. Int J SystBacteriol36:66–70Google Scholar
- Vaneechoutte M, De Beenhouwer H, Claeys G, Verschraegen G, De Rouck A, Paepe N, Elaichouni A, Portaels F (1993) Identification of mycobacterium species with amplified rDNA restriction analysis. J Clin Microbiol 31:2061–2065Google Scholar
- Wieland F, Lechner J, Sumper M (1982) The cell wall glycoprotein of halobacteium: structural, functional and biosynthetic aspects. Zbl Bakt Hyg J Abt Orig C3:161–170Google Scholar
- Winderickx J, Holsbeeks I, Lagatie O, Goits K, Thevelein J, Winde H (2003) In: yeast stress responses, topics in current genetics. Springer, Berlin, pp 307–386Google Scholar
- Yildiz E, Ozcan B, Caliskan M (2012) Isolation, characterization and phylogenetic analysis of Halophilic Archaea from a Salt Mine in Central Anatolia (Turkey). Pol J Microbiol 61:111Google Scholar