Isolation and characterization of the plasma membrane from slime-forming,encapsulated Streptococcus cremoris |
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| Institution: | 1. Department of Biotechnology, Dolphin (PG) Institute of Biomedical & Natural Sciences, Dehradun-248007, India;2. Algal Research and Bioenergy Lab, Department of Food Science and Technology, Graphic Era (Deemed to be University), Dehradun, Uttarakhand 248002, India;3. School of Engineering, University of Petroleum and Energy Studies, Dehradun, India;4. Department of Chemistry, Graphic Era (Deemed to be University), Dehradun, Uttarakhand 248002, India;5. Joint Institute for High Temperatures of the Russian Academy of Sciences, 13/2 Izhorskaya St, Moscow 125412, Russian Federation;6. Faculty of Applied Sciences and Biotechnology, Shoolini University, Solan 173229, HP, India;7. Peoples’ Friendship University of Russia (RUDN University), Moscow 117198, Russian Federation |
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| Abstract: | - 1.1. The plasma membrane of slime-forming, encapsulated Streptococcus cremoris from “viili” was isolated in hypotonie conditions in the presence of lysozyme (EC 3.2.1.17) using density gradient centrifugation as the last purification step.
- 2.2. The membrane yield was 15.8% of wet weight cells and the preparation contained 64.4% protein. 19.1% carbohydrate, 5.8% aminosugars, 5.1% RNA and 0.07% DNA.
- 3.3. Buffered 1% (w/v) Triton X-100 solubilized 33.6% of membrane proteins. The number of polypeptides detected by SDS-polyacrylamide gel electrophoresis was 59 when the membrane was isolated without a protease inhibitor and 44 in the presence of a protease inhibitor.
- 4.4. The molecular weights of the polypeptides varied from 13,500 to 100,000.
- 5.5. Ultrathin-layer electrofocusing analysis revealed the range of protein pi values to be between 3.50 and 5.85 concerning 77.3% of proteins and between pI 5.85 and 8.15 concerning 18.2% of proteins.
- 6.6. The isoelectric point of the only basic protein component was 9.3.
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