A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production
A rapid and economical method for the purification of phospholipase A1 (PLA1) from the extracellular medium of the ciliate Tetrahymena thermophila is presented. Essentially, the procedure, here designated as purification by selective interaction (PSI), entails the incubation of media containing PLA1...
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| Formato: | JOUR |
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_13645072_v86_n2_p226_Guberman |
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todo:paper_13645072_v86_n2_p226_Guberman2023-10-03T16:11:04Z A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production Guberman, A. Hartmann, M. Tiedtke, A. Florin-Christensen, J. Florin-Christensen, M. calcium chloride diethylaminoethyl cellulose liposome n,n dimethylformamide phospholipase a1 phospholipid article centrifugation enzyme activity enzyme purification ion exchange chromatography nonhuman polyacrylamide gel electrophoresis precipitation tetrahymena thermophila Animals Culture Media Liposomes Phospholipases A Tetrahymena thermophila Ciliophora Phaseolus (angiosperm) Protozoa Tetrahymena thermophila A rapid and economical method for the purification of phospholipase A1 (PLA1) from the extracellular medium of the ciliate Tetrahymena thermophila is presented. Essentially, the procedure, here designated as purification by selective interaction (PSI), entails the incubation of media containing PLA1 with liposomes made of soy bean phospholipids. The PLA1-lipid complexes are precipitated by the addition of CaCl2 and collected by centrifugation. Elution of the PLA1 is effected by treating the complexes with 40% dimethylformamide, a reversible inhibitor of this enzyme, which is easily removed by dialysis. In combination with DEAE cellulose ion exchange chromatography, PSI yielded homogeneous PLA1 preparations with a 14% recovery and a 416-fold increase in specific activity. This procedure, which can be completed within 1 day, may prove useful for the isolation of phospholipases from other sources. This practical method for the purification of a microbial PLA1 opens the way to large-scale production of these types of enzyme, which are not as yet commercially available. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_13645072_v86_n2_p226_Guberman |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
calcium chloride diethylaminoethyl cellulose liposome n,n dimethylformamide phospholipase a1 phospholipid article centrifugation enzyme activity enzyme purification ion exchange chromatography nonhuman polyacrylamide gel electrophoresis precipitation tetrahymena thermophila Animals Culture Media Liposomes Phospholipases A Tetrahymena thermophila Ciliophora Phaseolus (angiosperm) Protozoa Tetrahymena thermophila |
| spellingShingle |
calcium chloride diethylaminoethyl cellulose liposome n,n dimethylformamide phospholipase a1 phospholipid article centrifugation enzyme activity enzyme purification ion exchange chromatography nonhuman polyacrylamide gel electrophoresis precipitation tetrahymena thermophila Animals Culture Media Liposomes Phospholipases A Tetrahymena thermophila Ciliophora Phaseolus (angiosperm) Protozoa Tetrahymena thermophila Guberman, A. Hartmann, M. Tiedtke, A. Florin-Christensen, J. Florin-Christensen, M. A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production |
| topic_facet |
calcium chloride diethylaminoethyl cellulose liposome n,n dimethylformamide phospholipase a1 phospholipid article centrifugation enzyme activity enzyme purification ion exchange chromatography nonhuman polyacrylamide gel electrophoresis precipitation tetrahymena thermophila Animals Culture Media Liposomes Phospholipases A Tetrahymena thermophila Ciliophora Phaseolus (angiosperm) Protozoa Tetrahymena thermophila |
| description |
A rapid and economical method for the purification of phospholipase A1 (PLA1) from the extracellular medium of the ciliate Tetrahymena thermophila is presented. Essentially, the procedure, here designated as purification by selective interaction (PSI), entails the incubation of media containing PLA1 with liposomes made of soy bean phospholipids. The PLA1-lipid complexes are precipitated by the addition of CaCl2 and collected by centrifugation. Elution of the PLA1 is effected by treating the complexes with 40% dimethylformamide, a reversible inhibitor of this enzyme, which is easily removed by dialysis. In combination with DEAE cellulose ion exchange chromatography, PSI yielded homogeneous PLA1 preparations with a 14% recovery and a 416-fold increase in specific activity. This procedure, which can be completed within 1 day, may prove useful for the isolation of phospholipases from other sources. This practical method for the purification of a microbial PLA1 opens the way to large-scale production of these types of enzyme, which are not as yet commercially available. |
| format |
JOUR |
| author |
Guberman, A. Hartmann, M. Tiedtke, A. Florin-Christensen, J. Florin-Christensen, M. |
| author_facet |
Guberman, A. Hartmann, M. Tiedtke, A. Florin-Christensen, J. Florin-Christensen, M. |
| author_sort |
Guberman, A. |
| title |
A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production |
| title_short |
A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production |
| title_full |
A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production |
| title_fullStr |
A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production |
| title_full_unstemmed |
A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production |
| title_sort |
method for the preparation of tetrahymena thermophila phospholipase a1 suitable for large-scale production |
| url |
http://hdl.handle.net/20.500.12110/paper_13645072_v86_n2_p226_Guberman |
| work_keys_str_mv |
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1807316930484240384 |