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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Publicado:	1999
Materias:	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
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_13645072_v86_n2_p226_Guberman http://hdl.handle.net/20.500.12110/paper_13645072_v86_n2_p226_Guberman
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_13645072_v86_n2_p226_Guberman
record_format	dspace
spelling	paper:paper_13645072_v86_n2_p226_Guberman2023-06-08T16:11:47Z A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production 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. 1999 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_13645072_v86_n2_p226_Guberman 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 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.
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
publishDate	1999
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_13645072_v86_n2_p226_Guberman http://hdl.handle.net/20.500.12110/paper_13645072_v86_n2_p226_Guberman
_version_	1768545989692489728

A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production

Ejemplares similares