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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%.

acta chemica scandinavica/acta chemica scandinavica. b, organic chemistry and biochemistry/acta chemica scandinavica. a, physical and inorganic chemistry/acta chemica scandinavica. series b. organic chemistry and biochemistry/acta chemica scandinavica. series a, physical and inorganic chemistry

A Simple Purification Scheme Yielding Crystalline Phospholipase C from Bacillus cereus.