doi: 10.3891/acta.chem.scand.34b-0375.
A simple purification scheme yielding crystalline phospholipase C from Bacillus cereus
- PMID: 6781184
- DOI: 10.3891/acta.chem.scand.34b-0375
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A simple purification scheme yielding crystalline phospholipase C from Bacillus cereus
Acta Chem Scand B.
1980.
Abstract
A very simple and rather unusual purification scheme for phospholipase C from Bacillus cereus has been worked out. Air is bubbled vigorously through the bacterial culture and the foam collected. Liquified foam is centrifuged, dialyzed and heated at 74 degrees C for 5 min. After centrifugation, affinity chromatography is carried out on lipoprotein-Sepharose. The enzyme is then thermally denatured by exposure to 85 degrees C for 5 min and the precipitated material well washed and then renatured from solution in 4 M guanidinium chloride. The final enzyme preparations are electrophoretically homogeneous and easy to crystallize. The recovery of activity exceeds 80%.
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