Purification and characterisation of extracellular protease produced by Clostridium sp. from Schirmacher oasis,Antarctica |
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| Affiliation: | 1. Biotechnology Division, Defence R&D Establishment, Jhansi Road, Gwalior 474002, India;2. Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500007, India;3. Biochemistry Division, Defence R&D Establishment, Jhansi Road, Gwalior 474002, India;1. Icelandic Institute of Natural History, Iceland;2. Department of Geology, UiT The Arctic University of Norway, Norway;3. Centre for GeoGenetics, Natural History Museum of Denmark, University of Copenhagen, Denmark;4. Nordic Volcanological Center, Institute of Earth Sciences, University of Iceland, Iceland;5. Institute of Earth Sciences, University of Iceland, Iceland;6. The University Centre in Svalbard (UNIS), Norway;1. Birbal Sahni Institute of Palaeobotany, 53 University Road, Lucknow, 226007, India;2. Wadia Institute of Himalayan Geology, 33 General Mahadeo Singh Road, Dehradun, 248001, India;3. Waikhom Mani Girls'' College, Thoubal, 795138, Manipur, India;1. Department of Environmental Sciences, Norwegian University of Life Sciences, P.O. Box 5003, NO-1432 Ås, Norway;2. Department of Geology, Lund University, Sölvegatan 12, SE-223 62 Lund, Sweden;3. Faculty of Earth Sciences, University of Iceland, Sturlugata 7, IS-101 Reykjavik, Iceland;4. University Centre in Svalbard, UNIS, P.O. Box 156, NO-9171 Longyearbyen, Norway;1. Faculty of Earth Sciences, University of Iceland, Reykjavík, Iceland;2. INSTAAR and Department of Geological Sciences, University of Colorado Boulder, Boulder, CO, USA;1. DICA, Politecnico di Milano, P.zza L. da Vinci 32, I-20133 Milano, Italy;2. WSL Institute for Snow and Avalanche Research SLF, Flüelastr. 11, CH-7260 Davos Dorf, Switzerland |
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| Abstract: | One anaerobic, proteolytic bacterium isolated from Schirmacher oasis, Antarctica was characterised taxonomically. Based on morphology, biochemical characteristics and 16S rRNA sequence the isolate was identified as Clostridium species with closest similarity with Clostridium subterminale. Isolate was psychrotrophic forming maximum cell mass between 5 and 10 °C and produced extracellular protease. Growth was observed in the pH range of 6.5–8.5 with optimum at pH 8. Protease was purified 12.7-fold with a total yield of 26.2%. Effect of temperature, pH and salt concentration on enzyme activity were studied. Protease was found to be a serine-type metaloenzyme, which is active in a broad range of pH. It was moderately thermolabile and resistant to SDS. Enzyme kinetics revealed a tendency to decrease Km with increase in temperature for casein substrate. |
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