ISSN: 0973-7510

E-ISSN: 2581-690X

Ankita Sharma , Karuna Dhiman, Zeba Ansari and Poonam Shirkot
Department of Biotechnology, Dr Y S Parmar University of Horticulture and Forestry,
Nauni, Solan – 173 230 H.P., India.
J. Pure Appl. Microbiol., 2016, 10 (2): 1197-1210
© The Author(s). 2016
Received: 11/03/2016 | Accepted: 03/04/2016 | Published: 30/06/2016
Abstract

Four hot water springs viz., Manikaran, Vashisht, Khirganga and Tattapani were purposely selected for isolation of thermostable lipase producing bacterial isolates. The pH and temperature of the four thermal springs were  ranged from 4.0-6.0 and 51-105oC respectively. Isolated forty two thermophilic bacterial isolates, were described as putative thermostable lipase producers on the basis of their ability to form zone of clearance on tributyrin agar medium. Quantitative screening led to the selection of MBW2 bacterial isolate showing maximum thermostable lipase activity of 4.83U/ml after 24 hrs of incubation time at 60° C temperature, was selected for morphological, biochemical and molecular characterization. Genomic DNA  isolated from the selected MBW2 bacterial isolate was subjected to PCR amplification  followed by sequencing using universal primers for 16S rrna gene. In silico molecular analysis identified MBW2 bacterial isolate as Aneurinibacillus thermoaerophilus strain MBW2. Optimum culture conditions for growth and thermostable lipase activity of selected isolate were used for the enzyme production which was  purified to 3.08 fold with percent yield of 13.73% using ammonium sulphate precipitation technique, gel filteration chromatography and ion exchange chromatography technique.  The molecular weight of the  purified enzyme was found to be 42.5 kDa using SDS-PAGE.

Keywords

Thermostable lipase, Thermophiles, Hot water spring.

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© The Author(s) 2016. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.