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Proteinase affinity chromatography on bacitracin-Sepharose
Journal of Applied Biochemistry
|December 1, 1983
Summary
Bacitracin-Sepharose is an effective ligand for purifying various proteinases, including cysteine proteinases. This broad-spectrum ligand facilitates enzyme purification across major proteinase classes.
Area of Science:
- Biochemistry
- Enzymology
- Chromatography
Background:
- Proteinases are crucial enzymes involved in numerous biological processes.
- Affinity chromatography is a powerful technique for enzyme purification.
- Bacitracin, an antibiotic cyclopeptide, has shown potential as a general ligand.
Purpose of the Study:
- To evaluate bacitracin-Sepharose as a general ligand for affinity chromatography.
- To extend the application of bacitracin-Sepharose to cysteine proteinases.
- To demonstrate the broad applicability of bacitracin for proteinase purification.
Main Methods:
- Covalent immobilization of bacitracin onto Sepharose beads.
- Affinity chromatography using bacitracin-Sepharose.
- Purification of aspartyl, serine, metallo-, and cysteine proteinases from various sources.
Main Results:
- Bacitracin-Sepharose efficiently purified aspartyl, serine, and metalloproteinases with yields of 50-180%.
- New data confirm its efficacy for purifying cysteine proteinases like papain, bromelain, and ficin.
- Bacitracin effectively binds proteinases from all major classes, indicating broad specificity.
Conclusions:
- Bacitracin-Sepharose serves as a versatile and efficient general ligand for proteinase purification.
- The broad specificity of bacitracin as a weak proteinase inhibitor explains its effectiveness.
- This method offers a valuable tool for researchers studying diverse proteinases.