Purification and characterization of alkane solubilizing factor produced by Pseudomonas PG-1 |
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| Affiliation: | 1. Carbohydrate and Protein Biotechnology Laboratory, Department of Biotechnology, Punjabi University, Patiala, 147 002, Punjab, India;2. Chembiotech Laboratories, Advanced Science and Technology Institute, 5 The Croft, Buntsford Drive, Stoke Heath, Bromsgrove, Worcs, B60 4JE, UK;1. Department of Biological Sciences, P. D. Patel Institute of Applied Sciences, Charotar University of Science and Technology, Changa – 388 421, Anand, Gujarat, India;2. Multi-disciplinary Research Unit, Surat Municipal Institute of Medical Education & Research, Surat 395010, Gujarat, India;3. Gujarat Pollution Control Board, Gandhinagar, Gujarat 382 010, India;1. Department of Chemical Technologies, Expertise and Food Safety, Faculty of Integrated Technologies, Kherson National Technical University, st. Beryslavskoe shose, 24, 73008 Kherson, Ukraine;2. Department of Physics and Chemistry, University of Palermo, Viale delle scienze pad 17, 90128 Palermo, Italy;1. College of Marine Life Sciences, Ocean University of China, Yushan Road, No. 5, Qingdao, China;2. Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, 266003, China;1. Universidade Católica de Pernambuco (UNICAP), Rua do Príncipe, n. 526, Boa Vista, Recife Pernambuco CEP 50050-900, Brazil;2. Instituto Avançado de Tecnologia e Inovação (IATI), Rua Potyra, 31 - Prado, Recife Pernambuco CEP 50751-310, Brazil;3. Rede Nordeste de Biotecnologia (RENORBIO), Universidade Federal Rural de Pernambuco (UFRPE), Rua Dom Manuel de Medeiros, s/n, Dois Irmãos, CEP 52171-900 Recife, PE, Brazil;4. Faculty of Life and Health Sciences, School of Biomedical Sciences, University of Ulster, BT52 1SA Northern Ireland, UK;1. National Agricultural Research and Innovation Centre, Research Department of Field Crops, H-6726, Szeged, Alsókikötő sor 9, Hungary;2. Cereal Research Non-Profit Ltd., H-6726, Szeged, Alsókikötő sor 9, Hungary;3. Department of Physical Chemistry and Materials Science, University of Szeged, Aradi Vt. 1., H-6720, Szeged, Hungary;4. Laboratory of Proteomics Research, Institute of Biochemistry, Biological Research Centre of the Hungarian Academy of Sciences, H-6726, Szeged, Temesvári krt 62, Hungary;5. Institute of Organic Chemistry, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar tudósok körútja 2, H-1117, Budapest, Hungary |
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| Abstract: | The normal hexadecane emulsifying and solubilizing factor (PG-1 ESF C16) produced by Pseudomonas PG-1 during growth on n-hexadecane was isolated and purified. The factor was composed of protein, carbohydrate and lipid, which were largely undialyzable. Ca2+ was necessary for activation and heat stability of the factor. Particle size of the factor was less than 10 nm. All the protein along with 68–74% of the carbohydrate in the factor was obtained in a single protein peak by gel filtration chromatography using Biogel P-30. The isolated protein fraction showed a 1–5 fold increase in n-hexadecane solubilizing activity. The isolated protein was shown to be a homogeneous, monomeric protein with a molecular weight of approximately 11,000 daltons by SDS-PAGE. The protein and carbohydrate moieties in the isolate were separated by DEAE-cellulose chromatography. Neither purified protein nor carbohydrate showed n-hexadecane solubilizing activity separately, but when these were mixed full activity was restored. Hydrocarbon emulsifying activity was confined to the lipid fraction, which was isolated to the extent of 85% from the Biogel P-30 column by ethyl ether extraction. |
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