INVESTIGADORES
GUBERMAN Alejandra Sonia
artículos
Título:
A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production.
Autor/es:
GUBERMAN, A., HARTMANN, M., TIEDTKE, A., FLORIN-CHRISTENSEN J., FLORIN-CHRISTENSEN, M.
Revista:
JOURNAL OF APPLIED MICROBIOLOGY
Referencias:
Año: 1999 vol. 86 p. 226 - 230
ISSN:
1364-5072
Resumen:
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.