EVALUATION OF BIOTECHNOLOGICAL POTENTIAL OF THERMO – TOLERANT BACTERIA ISOLATED FROM MANIKARAN HOT SPRING, HIMACHAL PRADESH

EVALUATION OF BIOTECHNOLOGICAL POTENTIAL OF THERMO – TOLERANT BACTERIA ISOLATED FROM MANIKARAN HOT SPRING, HIMACHAL PRADESH

Sunita Devi1 and S. S. Kanwar2
1Department of Basic Sciences, College of Forestry, University of Horticulture and Forestry -Nauni, Solan (India)-173230
2Department of Microbiology, College of Basic Sciences, CSK, Himachal Pradesh Agricultural University, Palampur (India) – 176062
[Corresponding author E-mail1: sunitachamba@gmail.com ]
Received: 22-07-2015 Accepted: 14-08-2015
Abstract:
A total of 108 bacteria were isolated from three different types of samples viz. water, soil- sediment and biomat collected from upper Manikaran, lower Manikaran and Shangna sites selected at Manikaran hot spring, Himachal Pradesh. Screening of all the isolates for extracellular enzymes production revealed that lipase, cellulase, protease and amylase producing isolates were 84.26, 74.07, 43.52 and 42.59 per cent, respectively. None of the isolates was found positive for pectinolytic activity. Most of the isolates were multienzyme producers. Only seven efficient protease producers viz. S6, W15, B48, EpW3, BB4, EpSA1 and EpWB1, were examined quantitatively for protease production. Out of these, five isolates were found to exhibit optimum protease activity at 45°C while, two isolates at 55°C. Out of seven, five isolates were found to exhibit optimum protease activity at pH 7.0 while, one at pH 8.0 and one at 6.0. Analyses of thermostability profiles of crude proteases of W15 and EpSA1 showed that their Analyses of thermostability profiles of crude proteases of W15 and EpSA1 showed that their half lives at 60 and 65ºC were 30 and 15 min, respectively. Whereas, the half lives of B48, EpW3 and BB4 at 55 and 60ºC were 30 and 15 min, respectively. Half lives of crude protease of S6 at 70 and 75ºC were 30 and 15 min, respectively while, the half lives of crude protease of EpWB1 at 70, 90 and 100ºC were 60, 30 and 15 min, respectively. Thus, crude protease of EpWB1was found the most stable among these isolates. SDS-PAGE analysis of extracellular crude protease of EpWB1 revealed that it is made up of three polypeptide chains having different molecular weights i.e 20.0, 40.0 and 250 kDa. The isolate EpWB1 was identified as Thermoactinomyces vulgaris on the basis of 16S rRNA gene sequence analysis.

Keywords: bacteria, Manikaran hot spring, enzymes, SDS-PAGE

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