A method for the preparation of Tetrahymena thermophila phospholipase A1 suitable for large-scale production.
J Appl Microbiol
; 86(2): 226-30, 1999 Feb.
Article
in En
| MEDLINE
| ID: mdl-10063621
ABSTRACT
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.
Search on Google
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Phospholipases A
/
Tetrahymena thermophila
Limits:
Animals
Language:
En
Journal:
J Appl Microbiol
Journal subject:
MICROBIOLOGIA
Year:
1999
Document type:
Article
Affiliation country:
Argentina