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A simple purification scheme yielding crystalline phospholipase C from Bacillus cereus
Acta Chemica Scandinavica. Series B: Organic Chemistry and Biochemistry
|January 1, 1980
Summary
This study details a novel purification method for phospholipase C from Bacillus cereus, achieving high purity and activity recovery. The innovative technique utilizes foam collection and thermal treatments for efficient enzyme isolation.
Area of Science:
- Biochemistry
- Enzymology
- Microbiology
Background:
- Phospholipase C (PLC) is an important enzyme with various applications.
- Efficient purification of PLC from microbial sources like Bacillus cereus is crucial for research and industrial use.
Purpose of the Study:
- To develop a simple, unusual, and effective purification scheme for phospholipase C from Bacillus cereus.
- To obtain electrophoretically homogeneous and crystallizable enzyme preparations with high activity recovery.
Main Methods:
- Foam collection by air bubbling through bacterial culture.
- Sequential steps including centrifugation, dialysis, heat treatment (74°C, 5 min), affinity chromatography (lipoprotein-Sepharose), thermal denaturation (85°C, 5 min), washing, and renaturation (4 M guanidinium chloride).
Main Results:
- The developed purification scheme yielded electrophoretically homogeneous phospholipase C.
- The enzyme preparations were suitable for crystallization.
- Enzyme activity recovery exceeded 80%.
Conclusions:
- A highly efficient and straightforward purification protocol for Bacillus cereus phospholipase C has been established.
- This method offers a practical approach for obtaining pure and active enzyme for further studies and applications.