Abstract
Twenty-eight bacterial cultures, isolated from hot springs in Uttarakhand, were characterized with particular reference to their wide temperature and pH tolerance and production of enzymes in the thermophilic range. All the bacterial isolates were observed as Gram-positive or variable rods in varied arrangement. Bacterial isolates exhibited tolerance to a wide temperature range (20–80 °C), from mesophilic (+11° to +45 °C) to thermophilic (+46 ° to +75 °C); few almost reached the hyperthermophilic range (+76 °C). The isolates also tolerated a wide pH range (4–14) and moderate salt concentration. The optimum growth of the bacterial isolates was observed at 55 °C and 7 pH. Out of 28 isolates, 25 produced lipase, 25 amylase, 24 cellulase, 22 protease and 13 xylanase at 55 and 65 °C. Tolerance to a wide temperature and pH range and the production of enzymes in a thermophilic temperature range can be considered as indicators of ecological competence of these bacterial isolates for colonizing the high temperature environment. On the basis of 16S rDNA similarity, 20 bacterial isolates belonged to Bacillus licheniformis, five to Paenibacillus ehimensis and one each to Bacillus sonorensis, B. tequilensis, and Staphylococcus epidermidis. Besides variation in phenotypic characters, strains of B. licheniformis and P. ehimensis showed varying 16S rDNA similarity between 97–99 % and 95–99 %, respectively. Consideration of temperature preferences in classifying microorganisms on the basis of their minimum, maximum, and optimum growth requirements is also discussed. The study has ecological relevance in the context of colonization of high temperature environments by thermophilic bacteria.
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Aktuganov G, Melentjev A, Galimzianova N, Khalikova E, Korpela T, Susi P (2008) Wide-range antifungal antagonism of Paenibacillus ehimensis IB-X-b and its dependence on chitinase and beta-1,3-glucanase production. Can J Microbiol 54:577–587
Bhardwaj KN, Tiwari SC (2008) Geothermal energy resource utilization: perspectives of the Uttarakhand Himalaya. Curr Sci 95:846–850
Bhardwaj KN, Tiwari SC, Pandey A (2011) Conservation of thermophilic cyanobacterial diversity and physicochemical characteristics of thermal springs of tapoban geothermal field, Himalayan province (India). Nat Acad Sci Lett 34:77–82
Bouzas TM, Velázquez JB, Villa TG (2006) Industrial applications of hyperthermophilic enzymes: a review. Protein & Peptide Lett 13:645–651
Chien A, Edgar DB, Trela JM (1976) Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus. J Bacteriol 127:1550–1557
Cinti D, Pizzino L, Voltattorni N, Quattrocchi F, Walia V (2009) Geochemistry of thermal waters along fault segments in the beas and parvati valleys (north-west Himalaya, Himachal Pradesh) and in the sohna town (Haryana). Ind Geochem J 43:65–76
Dhakar K, Jain R, Tamta S, Pandey A (2014) Prolonged laccase production by a cold and pH tolerant strain of Penicillium pinophilum (MCC 1049) isolated from a low temperature environment. Enz Res (Article ID 120708)
Dwivedi V, Sangwan N, Nigam A, Garg N, Niharika N, Khurana P, Khurana JP, Lal R (2012) Draft genome sequence of Thermus sp. strain RL, isolated from a hot water spring located atop the Himalayan ranges at Manikaran, India. J Bacteriol 194:3534
Earl AM, Losick R, Kolter R (2008) Ecology and genomics of Bacillus subtilis. Trends Microbiol 16:269–275
Georlette D, Damien B, Blaise V, Depiereux E, Uversky VN, Gerday C, Feller G (2003) Structural and functional adaptations to extreme temperatures in psychrophilic, mesophilic, and thermophilic DNA Ligases. The J Biol Chem 278:37015–37023
Ghati A, Sarkar K, Paul G (2013) Isolation, characterization and molecular identification of esterolytic thermophilic bacteria from an Indian hot spring. Curr Res Microbiol Biotechnol 1:196–202
Ghosh D, Bal B, Kashyap VK, Pal S (2003) Molecular phylogenetic exploration of bacterial diversity in a Bakreshwar (India) hot spring and culture of Shewanella-related thermophiles. Appl Environl Microbiol 69(7):4332–4336
Haki GD, Rakshit SK (2003) Developments in industrially important thermostable enzymes: a review. Biores Technol 89:17–34
Karatas H, Uyar F, Tolan V, Baysal Z (2013) Optimization and enhanced production of α-amylase and protease by a newly isolated Bacillus licheniformis ZB-05 under solid-state fermentation. Ann Microbiol 63(1):45–52
Kumar B, Trivedi P, Mishra AK, Pandey A, Palni LMS (2004) Microbial diversity of soil from two hot springs in garhwal Himalaya. Microbiol Res 159:141–146
Kumar B, Pandey A, Palni LMS (2005) Extracellular amylase activity of saccharomycopsis fibuligera, a mycelial yeast isolated from a hot spring site in garhwal Himalayas. Ind J Microbiol 45:211–215
Kumar M, Yadav AN, Tiwari R, Prasanna R, Saxena AK (2013) Deciphering the diversity of culturable thermotolerant bacteria from Manikaran hot springs. Ann Microbiol. doi:10.1007/s13213–013–0709–7
Madslien EH, Olsen JS, Granum PE, Blatny JM (2012) Genotyping of B. licheniformis based on a novel multi-locus sequence typing (MLST) scheme. BMC Microbiol 12:230
Marsh CL, Larsen DH (1953) Characterization of some thermophilic bacteria from the hot springs of Yellowstone national park. J Bacteriol 65:193–197
Naing KW, Anees M, Kim SJ, Nam Y, Kim YC, Kim KY (2014) Characterization of antifungal activity of Paenibacillus ehimensis KWN38 against soilborne phytopathogenic fungi belonging to various taxonomic groups. Ann Microbiology 64(1):55–63
Nicholson WL, Munakata N, Horneck G, Melosh HJ, Setlow P (2000) Resistance of Bacillus endospores to extreme terrestrial and extraterrestrial environments. Microbiol Mol Biol Rev 64(3):548–572
Niehaus F, Bertoldo C, Kahler M, Antranikian G (1999) Extremophiles as a source of novel enzymes for industrial application. Appl Microbiol Biotechnol 51:711–729
Otto M (2009) Staphylococcus epidermidis- the ‘accidental’ pathogen. Nat Rev Microbiol 7:555–567
Palmisano MM, Nakamura LK, Duncan KE, Istock CA, Cohan FM (2001) Bacillus sonorensis sp. nov., a close relative of Bacillus licheniformis, isolated from soil in the sonoran desert, Arizona. Int J Syst Evol Microbiol 51:1671–1679
Pandey A, Palni LMS (1998) Isolation of Pseudomonas corrugata from Sikkim Himalaya. World J Microbiol Biotechnol 14:411–413
Pandey A, Palni LMS (2007) The rhizosphere effect in trees of the Indian central Himalaya with special reference to altitude. Appl Ecol Environ Res 5(1):93–102
Pandey A, Palni LMS, Coulomb N (1997) Antifungal activity of bacteria isolated from the rhizosphere of established tea bushes. Microbiol Res 152:105–112
Pandey A, Trivedi P, Kumar B, Palni LMS (2006) Characterization of a phosphate solubilizing and antagonistic strain of Pseudomonas putida (B0) isolated from a sub-alpine location in the Indian Central Himalaya. Curr Microbiol 53(2):102–107
Pandey A, Chaudhry S, Sharma A, Choudhary VS, Malviya MK, Chamoli S, Rinu K, Trivedi P, Palni LMS (2011) Recovery of Bacillus and Pseudomonas spp from the ‘fired plots’ under shifting cultivation in northeast India. Curr Microbiol 62:273–280
Pandey A, Dhakar K, Sati P, Sharma A, Kumar B, Palni LMS (2013) Geobacillus stearothermophilus (GBPI_16): a resilient hyperthermophile isolated from an autoclaved sediment sample. Proc Nat Acad Sci India (Section B). doi:10.1007/s40011–013–0210-x
Pennisi E (1997) In industry, extremophiles begin to make their mark. Science 276:705–706
Pesciaroli C, Cupini F, Selbmann L, Barghini P, Fenice M (2012) Temperature preferences of bacteria isolated from seawater collected in Kandalaksha bay. White Sea, Russia Polar Biol 35:435–445
Purcell D, Sompong U, Lau CY, Barraclough TG, Peerapornpisal Y, Pointing SB (2007) The effects of temperature, pH and sulphide on community structure of hyperthermophilic streamers in hot springs of northern Thailand. FEMS Microbiol Ecol 60:456–466
Rinu K, Pandey A (2010) Temperature dependent phosphate solubilization by cold and pH tolerant species of aspergillus isolated from Himalayan soil. Mycoscience 51(4):263–271
Rinu K, Pandey A (2011) Slow and steady phosphate solubilization by a psychrotolerant strain of Paecilomyces hepiali (MTCC 9621). World J Microbiol Biotechnol 27(5):1055–1062
Rothschild LJ, Mancinelli RL (2001) Life in extreme environments. Nature 409:1092–1101
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Sharma A, Pandey A, Shouche Y, Kumar B, Kulkarni G (2009) Characterization and identification of Geobacillus spp. Isolated from soldhar hot spring site of garhwal Himalaya, India. J Basic Microbiol 48:187–194
Takacs-Vesbach C, Mitchell K, Jackson-Weaver O, Reysenbach AL (2008) Volcanic calderas delineate biogeographic provinces among Yellowstone thermophiles. Environ Microbiol 10:1681–1689
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 24:4876–4882
Tobler JD, Benning LG (2011) Bacterial diversity in five Icelandic geothermal waters: temperature and sinter growth rate effects. Extremophiles 15:473–485
Trivedi P, Kumar B, Pandey A (2006) Conservation of soil microbial diversity associated with two hot springs in Uttaranchal Himalaya. Nat Acad Sci Lett 29:185–188
Valverde A, Tuffin M, Cowan DA (2012) Biogeography of bacterial communities in hot springs: a focus on the actinobacteria. Extremophiles 16:669–679
van Dijl JM, Hecker M (2013) Bacillus subtilis: from soil bacterium to supersecreting cell factory. Microb Cell Factories 12:3
Whitaker RJ, Grogan DW, Taylor JW (2003) Geographical barriers isolate endemic population of hyperthermophilic archaea. Science 301:976–978
Wiegel J (1990) Temperature spans for growth: hypothesis and discussion. FEMS Microbiol Lett 75:155–169
Wolferen M, Ajon M, Driessen AJM, Albers S (2013) How hyperthermophiles adapt to change their lives: DNA exchange in extreme conditions. Extremophiles 17:545–563
Yates C, Gilling MR, Davison AD, Altavilla N, Veal DA (1997) PCR amplification of crude microbial DNA extracted from soil. Lett Appl Microbiol 25:303–307
Zheng H, Wu H (2010) Gene-centric association analysis for the correlation between the guanine cytosine content levels and temperature range conditions of prokaryotic species. BMC Bioinformatics 10 pages
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The director of GBPIHED is acknowledged for extending use of the facilities. The Ministry of Environment and Forests, Govt. of India, New Delhi, is acknowledged for financial support.
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Pandey, A., Dhakar, K., Sharma, A. et al. Thermophilic bacteria that tolerate a wide temperature and pH range colonize the Soldhar (95 °C) and Ringigad (80 °C) hot springs of Uttarakhand, India. Ann Microbiol 65, 809–816 (2015). https://doi.org/10.1007/s13213-014-0921-0
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DOI: https://doi.org/10.1007/s13213-014-0921-0